Note: Descriptions are shown in the official language in which they were submitted.
1
TRIAZOLE FORMULATIONS
Related Applications
This application claims priority to United States Provisional Patent
Application serial number
61/758,914 filed January 31, 2013 and to United States Provisional Patent
Application serial number
61/763,127 filed on February 11, 2013.
Background
Triazole fungicides are used on a wide variety of plants in agriculture
including field crops,
fruit trees, small fruit, vegetables and turf. Triazoles are used against a
variety of fungi, including but
not limited to powdery mildews, rusts and leaf-spotting fungi. Exemplary
fungicides include but are
not limited to difenoconazole, fenbuconazole, myclobutanil, propiconazole,
tebuconazole,
tetraconazole, triticonazole and epiconazole.
Triazoles are believed to inhibit enzymes used in the production of cell
membranes and cells
walls. Their use results in abnormal fungi growth and death. Each triazole
functions in a different
part of the cell membrane/wall formation process; therefore, there is wide
variability in the activity
spectra amongst triazoles and target fungi.
Triazoles can be applied as a preventative fungicide and also as a curative
fungicide. In
curative treatments, the fungicide is traditionally best applied before spore
formation as triazoles
are not effective in inhibiting spore formation. Triazole pesticides exhibit
some systemic activity
(e.g., within a leaf) and this activity varies across the class of compounds.
Some triazoles are
systemic within local structures, and are not transported from one part of a
plant to another, while
other triazole compounds are more widely transported through the plant.
Triazoles are currently formulated into various usable forms such as
emulsifiable
concentrates (ECs), liquid concentrates (SL), and other forms that use
petroleum or non-petroleum
based solvents along with anionic or non-ionic emulsifiers and stabilizers to
compensate for low
water solubility, low soil motility and other drawbacks of triazoles based on
their chemical
properties. Furthermore, triazoles also vary in their photolytic stability
under natural environmental
conditions; therefore formulations often developed to compensate and reduce
the susceptibility to
chemical degradation before and after the formulation has been applied to a
crop. There remains a
Date Recue/Date Received 2020-06-05
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
2
need for improved formulations that reduce the dependence on additives and
formulants, yet also
prove as effective as current formulations.
Furthermore, because triazoles have a very specific mode of action, targeted
fungi can
become resistant. Different formulation techniques have therefore been
developed in an attempt
to address these deficiencies. An ideal formulation would have adequate
loading of the active
ingredient, be non-odorous, non-caking, non-foaming, stable under extreme
conditions for extended
periods of time, disperse rapidly upon addition to a spray tank, be compatible
with a range of
secondary additives and other agricultural products (fertilizer, pesticide,
herbicide and other
formulations) added to a spray tank, pourable or flowable, and, for solid
formulations, be non-dusty
(for solid formulations), and have sufficient/superior rainfast properties
after application.
Summary of the Invention
The present disclosure provides formulations of triazole compounds including
nanoparticles
of polymer-associated triazole compounds with various formulating agents. The
present disclosure
also provides methods of producing and using these formulations.
In various embodiments, the present disclosure presents formulations including
a
nanoparticle including a polymer-associated triazole compound with an average
diameter of
between about 1 nm and about 500 nm; and the polymer is a polyelectrolyte and
a dispersant or a
wetting agent.
In some embodiments, the nanoparticle has a diameter of between about 1 nm and
about
100 nm. In some embodiments, the nanoparticle has a diameter of between about
1 nm and about
20 nm.
In some embodiments, the formulation includes a plurality of nanoparticles,
wherein the
nanoparticles are in an aggregate and the aggregate has a diameter of between
about 10 nm and
about 5000 nm. In some embodiments, the formulation includes a plurality of
nanoparticles,
wherein the nanoparticles are in an aggregate and the aggregate has a diameter
of between about
100 nm and about 2500 nm. In some embodiments, the formulation includes a
plurality of
nanoparticles, wherein the nanoparticles are in an aggregate and the aggregate
has a diameter of
between about 100 nm and about 1000 nm. In some embodiments, the formulation
includes a
plurality of nanoparticles, wherein the nanoparticles are in an aggregate and
the aggregate has a
diameter of between about 100 nm and about 300 nm.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
3
In some embodiments, the ratio of triazole compound to polymer within the
nanoparticles is
between about 10:1 and about 1:10. In some embodiments, the ratio of triazole
compound to
polymer within the nanoparticles is between about 5:1 and about 1:5. In some
embodiments, the
ratio of triazole compound to polymer within the nanoparticles is between
about 2:1 and about 1:2.
In some embodiments, the ratio of triazole compound to polymer within the
nanoparticles is about
1:3. In some embodiments, the ratio of triazole compound to polymer within the
nanoparticles is
about 3:2. In some embodiments, the ratio of triazole compound to polymer
within the
nanoparticles is about 4:1. In some embodiments, the ratio of triazole
compound to polymer within
the nanoparticles is about 2:1. In some embodiments, the ratio of triazole
compound to polymer
within the nanoparticles is about 1:1. In some embodiments, the triazole
compound is
difenoconazole.
In some embodiments, the polymer is selected from the group consisting of
poly(methacrylic
acid co-ethyl acrylate); poly(nnethacrylic acid-co-styrene); poly(nnethacrylic
acid-co-
butylnnethacrylate); poly[acrylic acid-co-poly(ethylene glycol) methyl ether
methacrylate]; poly(n-
butylnnethacrylcate-co-nnethacrylic acid) and poly(acrylic acid-co-styrene. In
some embodiments,
the polymer is a honnopolynner. In some embodiments, the polymer is a
copolymer. In some
embodiments, the polymer is a random copolymer.
In some embodiments, the dispersant and/or wetting agent is selected from the
group
consisting of lignosulfonates, organosilicones, methylated or ethylated seed
oils, ethoxylates,
sulfonates, sulfates and combinations thereof. In some embodiments, the
dispersant and/or
wetting agent is sodium lignosulfonate. In some embodiments, the dispersant
and/or wetting agent
is a tristyrylphenol ethoxylate. In some embodiments, the wetting agent and
the dispersant are the
same compound. In some embodiments, the wetting agent and the dispersant are
different
compounds.
In some embodiments, the formulation excludes any wetting agent. In some
embodiments,
the formulation excludes any dispersant. In some embodiments, the wetting
agent is less than
about 30 weight % of the formulation. In some embodiments, the wetting agent
is less than about 5
weight % of the formulation. In some embodiments, the dispersant is less than
about 30 weight %
of the formulation. In some embodiments, the dispersant is less than about 5
weight % of the
formulation. In some embodiments, the formulation is in the form of a high
solids liquid suspension
or a suspension concentrate.
In some embodiments, the formulation includes between about 0.05 weight % and
about 5
weight % of a thickener. In some embodiments, the thickener is less than about
1 weight % of the
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
4
formulation. In some embodiments, the thickener is less than about 0.5 weight
% of the
formulation. In some embodiments, the thickener is less than about 0.1 weight
% of the
formulation. In some embodiments, the thickener is selected from the group
consisting of guar
gum; locust bean gum; xanthan gum; carrageenan; alginates; methyl cellulose;
sodium
carboxymethyl cellulose; hydroxyethyl cellulose; modified starches;
polysaccharides and other
modified polysaccharides; polyvinyl alcohol; glycerol alkyd, fumed silica and
combinations thereof.
In some embodiments, the formulation includes between about 0.01 weight % and
about
0.2 weight % of a preservative. In some embodiments, the preservative is less
than about 0.1 weight
% of the formulation. In some embodiments, the preservative is less than about
0.05 weight % of
the formulation. In some embodiments, the preservative is selected from the
group consisting of
tocopherol, ascorbyl palnnitate, propyl gallate, butylated hydroxyanisole
(BHA), butylated
hydroxytoluene (BHT), propionic acid and its sodium salt; sorbic acid and its
sodium or potassium
salts; benzoic acid and its sodium salt; p-hydroxy benzoic acid sodium salt;
methyl p-hydroxy
benzoate; 1,2-benzisothiazalin-3-one, and combinations thereof.
In some embodiments, the formulation includes between about 0.05 weight % and
about 10
weight % of an anti-freezing agent. In some embodiments, the anti-freezing
agent is less than about
weight % of the formulation. In some embodiments, the anti-freezing agent is
less than about 1
weight % of the formulation. In some embodiments, the anti-freezing agent is
selected from the
group consisting of ethylene glycol; propylene glycol; urea and combinations
thereof.
In some embodiments, the polymer-associated triazole compound is less than
about 80
weight % of the formulation. In some embodiments, the polymer-associated
triazole compound is
between about 20 weight % and about 80 weight % of the formulation. In some
embodiments, the
polymer-associated triazole compound is between about 20 weight % and about 50
weight % of the
formulation. In some embodiments, the polymer-associated triazole compound is
between about 5
weight % and about 40 weight % of the formulation.
In some embodiments, the triazole compound is selected from the groups
consisting of
difenoconazole, fenbuconazole, nnyclobutanil, propiconazole, tebuconazole,
tetraconazole,
triticonazole and epiconazole.
In some embodiments, the formulation includes an inert filler. In some
embodiments, the
inert filler makes up less than about 90 weight % of the formulation. In some
embodiments, the
inert filler makes up less than about 40 weight % of the formulation. In some
embodiments, the
inert filler makes up less than about 5 weight % of the formulation. In some
embodiments, the inert
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
filler is selected from the group consisting of saccharides, celluloses,
starches, carbohydrates,
vegetable oils, protein inert fillers, polymers and combinations thereof.
In some embodiments, the formulation includes between about 1 weight % and
about 20
weight % of a disintegrant. In some embodiments, the formulation includes
between about 0.05
weight % and about 3 weight % of an anti-caking agent. In some embodiments,
the anti-caking agent
is less than about 1 weight % of the formulation. In some embodiments, the
formulation includes
between about 0.05 weight % and about 5 weight % of an anti-foaming agent. In
some
embodiments, the anti-foaming agent is less than about 1 weight % of the
formulation.
In some embodiments, the formulation includes between about 1 weight % and
about 20
weight % of a non-ionic surfactant. In some embodiments, the non-ionic
surfactant is less than
about 1 weight % of the formulation.
In some embodiments, the formulation is diluted so that the concentration of
the polymer-
associated triazole compound is between about 0.1 to about 1000 ppm. In some
embodiments, the
formulation is diluted so that the concentration of the polymer-associated
triazole compound is
between about 10 to about 500 ppm. In some embodiments, the formulation also
includes a
strobilurin fungicide.
In various aspects, the present disclosure describes a method of using any of
the
formulations described herein by applying the formulation to a plant. In some
embodiments, the
formulation is applied to one part of a plant and the triazole translocates to
an unapplied part of the
plant. In some embodiments, the unapplied part of the plant comprises new
plant growth since the
application.
In various aspects, the present disclosure describes a method of inoculating a
plant with a
triazole against fungi by applying any of the formulations described herein.
In various aspects, the
present disclosure provides a method of treating a fungal infection of a plant
with a triazole by
applying any of the formulations described herein, to the plant. In various
aspects, the present
disclosure describes a method of increasing a plant's fungus resistance by
applying any of the
formulations described herein, to the plant.
In some embodiments, the plant to which the formulation is applied is selected
from the
classes fabaceaae, brassicaceae, rosaceae, solanaceae, convolvulaceae,
poaceae, annaranthaceae,
laminaceae and apiaceae. In some embodiments, the plant to which the
formulation is applied is
selected from oil crops, cereals, pasture, turf, ornamentals, fruit, legume
vegetables, bulb
vegetables, cole crops, tobacco, soybeans, cotton, sweet corn, field corn,
potatoes and greenhouse
crops. In some embodiments, the fungi targeted is selected from the classes
asconnycota,
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
6
basidiomycota, deuteromycota, blastocladiomycota, chytridiomycota,
glomeromycota and
combinations thereof.
In various aspects, the present invention is a formulation including a
nanoparticle comprising
a polymer-associated triazole compound with an average diameter of between
about 1 nnn and
about 500 nnn; wherein the polymer is a polyelectrolyte, a taurate dispersant,
a polycarboxylate salt
wetting agent, an anti-foaming agent, a preservative, and water.
In some embodiments, the triazole compound constitutes between about 5 and
about 30
percent by weight of the formulation. In some embodiments, the ratio of the
weight percent of the
triazole compound to the weight percent of the nanoparticles is between about
1:1 to 6:1. In some
embodiments, the formulation also includes a thickener.
In some embodiments, the formulation also includes an anti-freeze agent. In
some
embodiments, the formulation also includes an olefin sulfonate salt
surfactant. In some
embodiments, the formulation also includes a block copolymer surfactant. In
some embodiments,
the formulation also includes an additional pesticidal compound. In some
embodiments, the
additional pesticidal compound is a fungicide. In some embodiments, the
fungicide is a strobilurin.
In some embodiments, the polyelectrolyte polymer is a poly(nnethacrylic acid-
co-styrene) polymer.
In some embodiments, the taurate dispersant constitutes between about 0.5
weight percent
and about 5 weight percent of the formulation. In some embodiments, the
polycarboxylate salt
wetting agent constitutes between about 0.5 weight percent and about 5 weight
percent of the
formulation. In some embodiments, the anti-foaming agent constitutes between
about 0.1 weight
percent and about 1 weight percent of the formulation. In some embodiments the
preservative
constitutes between about 0.01 weight percent and about 0.1 weight percent of
the formulation. In
some embodiments, the thickener constitutes between about 0.05 weight percent
and about 2
weight percent of the formulation.
In some embodiments, the anti-freeze agent constitutes between about 1 weight
percent
and about 10 weight percent of the formulation. In some embodiments, the
olefin sulfonate salt
surfactant constitutes between about 0.5 weight percent and about 5 weight
percent of the
formulation. In some embodiments, the block copolymer surfactant constitutes
between about 0.5
weight percent and about 5 weight percent of the formulation. In some
embodiments, the
additional pesticide constitutes between about 5 weight percent and about 30
weight percent of the
formulation.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
7
Brief Description of the Drawings
Figure 1 is a graph illustrating the percent of disease controlled on a
disease incidence basis
over the course of several applications for two fungicide formulations,
InspireTM, a commercially
available formulation, and a nanoparticle formulation as described in Example
1. The disease is
Black Spot on cabbages as described in Example 3 and the disease control
figures are over the course
of second and third applications of the formulations.
Figure 2 is a graph illustrating the percent of disease controlled (based on
disease incidence)
over the course of two applications of two different fungicide formulations, a
commercially available
formulation and a formulation as described below in Example 1. Rates of
control were averaged for
three different application rates. The disease is powdery mildew (pathogen:
Golovinomyces
cichoracearu) on cantaloupe plants, as described in Example 4.
Figure 3 is a graph illustrating percent of disease controlled (based on
disease incidence) for
different application rates of two fungicide formulations at different
application rates of active
ingredient 18 days after a third treatment. The disease, crop treated and
application protocol are all
described in Example 4.
Figure 4 is a graph illustrating the percent of disease (based on disease
severity) controlled
14 days after application of two different fungicide formulations, a
commercially available
formulation and a formulation as described below in Example 1. Three different
application rates for
each formulation were evaluated. The disease is powdery mildew (pathogen:
Podosphaera xanthii)
on squash plants, as also described in Example 4.
Figures 5A & 5B illustrate rates of disease control, based on disease
incidence and severity,
respectively, for treatment of powdery mildew on squash plants as described in
Example 4.
Evaluations in these figures were performed 12 days after a second
application.
Figure 6 illustrate rates of disease control for two different formulations at
various
application rates and with an additional non-ionic surfactant added in
dilution step. The disease is
Peanut Leaf Spot on peanut plant as described in Example 5.
Figure 7 is a graph illustrating expected yield of peanut plants infected with
Peanut Leaf Spot
for various treatments.
Figure 8 is a graph illustrating percent of disease controlled (based on
disease incidence) for
different application rates of two fungicide formulations (Inspire and the
formulation described in
Example 1) at different application rates of active ingredient 14 days after
treatment. The disease
was Frog-Eye Leaf Spot on soybean plants as described in Example 6.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
8
Figure 9 is a graph illustrating different yields based on different
treatments of soybean
plants infected with Frog-Eye Leaf Spot as described in Example 6.
Figure 10 is a graph illustrating percent of disease controlled (based on
disease severity) for
different application rates of two fungicide formulations (Inspire and the
formulation described in
Example 1) at different application rates of active ingredient 6 days after
treatment. The disease
was Early Blight on tomato plants as described in Example 7.
Figure 11 is a graph illustrating percent of disease controlled (based on
disease severity) for
different application rates (averaged together) of two fungicide formulations
(InspireTM and the
formulation described in Example 2) at different points in a treatment regime.
The disease was
powdery mildew on zucchini plants as described in Example 8.
Figure 12 is a graph illustrating percent of disease controlled (based on
disease severity) for
different application rates (averaged together) of two fungicide formulations
(lnspireTM and the
formulation described in Example 2) at different points in a treatment
regimen. The disease was
powdery mildew on zucchini as described in Example 8.
Figure 13 is a graph illustrating disease index at various time points during
a treatment
regimen for three different fungicide formulations applied to the plants
(bananas) at a rate of 667
ppm (a commercial emulsifiable concentrate (labelled "Syngenta EC")), the
formulation described in
Example 2 ("VCP-05"), and a proprietary oil-in-water formulation ("Hainan
Zheng Ye EW")) at
different points in a treatment regimen. The disease was Sigatoka Leaf Spot on
banana plants. The
treatment program and evaluation methods are described in Example 9.
Figure 14 is a graph illustrating percent of disease controlled (based on
disease index shown
in Figure 13) for different application rates (250 ppm, 417 ppm and 667 ppm)
of the three fungicide
formulations described above in Figure 13 upon completion of the treatment
program. The disease,
crop treated, treatment program, and evaluation methods are all described in
Example 9.
Figure 15 is a graph illustrating percent of disease level for two different
difenoconazole
formulations (InspireTM, and a formulation prepared according to Example 2).
Disease level for an
untreated control is also shown on Figure 15. Disease level for each
formulation was averaged
between two different application rates (75 g active ingredient/ha and 125 g
active ingredient/ha).
Full details of the field test are described in Example 10.
Figure 16 is a graph illustrating percent of disease level for two different
fungicide
formulations (MuscleTm, a commercially available emulsifiable concentrate of
tebuconazole, and a
formulation prepared according to Example 2). The difenoconazole formulation
of Example 2 was
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
9
applied at two different application rates (75 g a.i./ha and 125 g a.i./ha).
Full details of the field test
are described in Example 10.
Figure 17 shows peanut yield rates for an entire growing season in which test
plots were
treated with various fungicides (e.g, difenoconazole (VCP-05), chlorothalonil
(EchoTm), chlorothalonil
mixed with prothioconazole (EchoTm/ProvostIm)) and different tank-mix, non-
ionic surfactants
(SilwetTM L-77 & InduceTm). Field test methods are described in Example 10.
Figure 18 is a graph showing disease level (measured by percent of row feet of
crop
infected) for two difenoconazole formulations a various application rates and,
in the case of the VCP-
05 formulation, with different tank-mixed non-ionic-surfactants. The disease
targeted was white
mold on peanuts and the field trial is described in Example 11.
Figure 19 shows a graph of peanut yield rates for an entire growing season in
which test
plots were treated with various fungicides (e.g, difenoconazole (VCP-05),
chlorothalonil (BravoTm),
chlorothalonil mixed with prothioconazole (BravoTm/ProvostTm)) and different
tank-mix, non-ionic
surfactants (SilwetTM L-77 & InduceTm). Field test methods are described in
Example 11.
Figure 20 is a graph showing disease control rates for a difenoconazole
formulation, VCP-05,
applied to treat dollar spot on creeping bentgrass. The disease control rates
for three different
application rates (0.25, 0.5 and 1.0 fluid oz. of formulation per 1000 ft2
treated area). Field test
procedures and evaluation methods are described in Example 12.
Figure 21 is a graph showing disease control rates for two
difenoconazole/azoxystrobin
mixture formulations. The first mixture was VCP-05 was mixed with HeritageTm,
a commercially
available azoxystrobin formulation. The second mixture was BriskwayTM, a
commercially available
formulation containing difenoconazole and azoxystrobin. The formulations were
applied to treat
dollar spot on creeping bentgrass. Field test procedures and evaluation
methods are described in
Example 13.
Figure 22 is a graph showing disease control rates for a difenoconazole
formulation, VCP-05,
applied to treat anthracnose on annual bluegrass. The disease control rates
for three different
application rates (0.25, 0.5 and 1.0 fluid oz. of formulation per 1000 ft2
treated area). Field test
procedures and evaluation methods are described in Example 14.
Definitions
As used herein, the term "inoculation" refers to a method used to administer
or apply a
formulation of the present disclosure to a target area of a plant or fungus.
The inoculation method
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
can be, but is not limited to, aerosol spray, pressure spray, direct watering,
and dipping. Target
areas of a plant could include, but are not limited to, the leaves, roots,
stems, buds, flowers, fruit,
and seed. Target areas of the fungus could include, but are not limited to,
the hyphae and
mycelium, inoculating reproductive spores (conidia or ascospores) and the
haustoria. Inoculation
can include a method wherein a plant is treated in one area (e.g., the root
zone or foliage) and
another area of the plant becomes protected (e.g., foliage when applied in the
root zone or new
growth when applied to foliage). Inoculation can also include a method wherein
a plant is treated in
one area (e.g., the foliar surface) and fungal infection in the interior of
the plant is cured.
As used herein, the term "wettable granule" also referred to herein as "WG"õ
and "soluble
granule" refers to a solid granular formulation that is prepared by a
granulation process and that
contains nanoparticles of polymer-associated active ingredient, (includes
potentially aggregates of
the same), a wetting agent and/or a dispersant, and optionally an inert
filler. Wettable granules can
be stored as a formulation, and can be provided to the market and/or end user
without further
processing. In some embodiments, they can be placed in a water-soluble bag for
ease of use by the
end user. In most practical applications, wettable granules are prepared for
application by the end
user. The wettable granules are mixed with water in the end user's spray tank
to the proper dilution
for the particular application. Dilution can vary by crop, fungus, time of
year, geography, local
regulations, and intensity of infestation among other factors. Once properly
diluted, the solution
can be applied by e.g., spraying.
As used herein, the term "wettable powder" also referred to herein as "WP",
"water
dispersible powder" and "soluble powder", refers to a solid powdered
formulation that contains
nanoparticles of polymer-associated active ingredient (includes potentially
aggregates of the same), and
optionally one or more of a dispersant, a wetting agent, and an inert filler.
Wettable powders can be
stored as a formulation, and can be provided to the market and/or end user
without further
processing. In some embodiments, they can be placed in a water-soluble bag for
ease of use by the
end user. In practical applications, a wettable powder is prepared for
application by the end user.
The wettable powder is mixed with water in the end user's spray tank to the
proper dilution for the
particular application. Dilution can vary by crop, fungus, time of year,
geography, local regulations,
and intensity of infestation among other factors. Once properly diluted, the
solution can be applied
by e.g., spraying.
As used herein, the term "high solids liquid suspension" also referred to
herein as "HSLS"
refers to a liquid formulation, similar to a suspension concentrate, that
contains nanoparticles of
polymer nanoparticles associated with active ingredient (includes potentially
aggregates of the
same), a wetting agent and/or a dispersant, an anti-freezing agent, optionally
an anti-settling agent
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
11
or thickener, optionally a preservative, and water. High solids liquid
suspensions can be stored as a
formulation, and can be provided to the market and/or end user without further
processing. In
most practical applications, high solids liquid suspensions are prepared for
application by the end
user. The high solids liquid suspensions are mixed with water in the end
user's spray tank to the
proper dilution for the particular application. Dilution can vary by crop,
fungus, time of year,
geography, local regulations, and intensity of infestation among other
factors. Once properly
diluted, the solution can be applied by e.g., spraying.
Description of Various Embodiments of the Invention
Triazoles represent a very important class of fungicide globally. Triazoles
are used in
agriculture to protect crops such as cereals, field crops, fruits, tree nuts,
vegetables, turfgrass and
ornamentals because of their broad spectrum activity as well as (to varying
degrees) their activity
against all three major groups of plant pathogenic fungi: Ascomycetes,
Basidionnycetes, and
Deuteronnycetes. Triazoles also have found use outside agricultural
applications, such as human and
veterinary antifungal formulations.
Solubility
Triazoles as a class are typically poorly soluble in water, generally with
solubilities in the
parts per million range, or lower. Triazole solubilities are generally higher
in organic solvents (e.g.,
hexane, ethanol, dichloronnethane). See Table 1 below for a list of typical
triazoles, their solubilities
in several solvents, octanol-water partition coefficients and their melting
points. (Data via the
Pesticide Properties Database)
Table 1: Solubility of exemplary triazoles in common solvents, octanol-water
partition coefficients
and melting points
Triazole Solubility mg/L (solvent & conditions) Kow Melting
Point ( C)
Difenoconazole 15.0 mg/L (water at 20 C)
log P: 4.36 82.5
3400 mg/L (hexane at 20 C)
330000 mg/L (ethanol at 20 C)
Epoxiconazole 7.1 mg/L (water at 20 C) log P: 3.3 136.7
28800 mg/L (ethanol at 20 C)
Tebuconazole 36 mg/L (water at 20 C) log P: 3.7 105
80 mg/L (hexane at 20 C) (decomposes
2000000 mg/L (dichloromethane at 20 C) at 350)
Triticonazole 9.3 mg/L (water at 20 C) log P: 3.29 137
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
12
120 nng/L (hexane at 20 C)
18200 mg/L (methanol at 20 C)
Propiconazole 150 nng/L (water at 20 C) log P: 3.72 -23
1585 nng/L (heptane at 20 C) (decomposes
at 355)
Myclobutanil 132 nng/L (water at 20 C) log P: 2.89 70.9
1020 nng/L (heptane at 20 C)
250000 nng/L (methanol and acetone, both at
20 C)
Cyproconazole 93 nng/L (water at 20 C) log P: 3.09 106.5
1300 nng/L (hexane at 20 C)
Tetrazonazole 156.6 nng/L (water at 20 C) log P: 3.56 -29.2
300000 nng/L (xylene, acetone, ethyl acetate, (degrades at
all at C) 235)
Improvements in triazole solubility are desirable in order to improve
formulation processes,
simplify formulations, reduce the environmental consequences in fungicide
application and improve
fungicide efficacy.
Photolysis/Stability
Triazoles vary in their degradation rates upon exposure to sunlight and
demonstrate a range
of half-lives as listed in Table 2.
Table 2: Photolytic stability of some Triazoles
Triazole Photolytic Stability
Difenoconazole Stable at pH 7
Epoxiconazole DT50: 52d (aqueous photolysis at pH 7)
Tebuconazole Stable, no significant photolytic degradation
Triticonazole DT50: 3.1d (aqueous photolysis at pH 7)
Propiconazole Stable at pH 7
Myclobutanil DT50: 15d (aqueous photolysis at pH 7)
Cyproconazole DT 50: 40d (aqueous photolysis at pH 7)
Tetraconazole DT50: 217d (stable at pH 7)
Due to the tendency of some triazoles to degrade upon exposure to sunlight,
some crop
protection formulations of triazoles employ a UV blocker such as zinc, tin or
iron oxides as well as
organic UV blockers (e.g., 1,2-dihydroxybenzophenone). The use of UV-blockers
in formulation can
present additional complications in formulating, application and use. For
example, the UV-blocker
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
13
is an additional component that needs to be soluble or at least dispersible in
the media or matrix of
the product. It is therefore desirable to produce formulations that do not
require a UV-blocker.
Fungicide Resistance
Triazoles are site specific fungicides and inhibit one specific enzyme, C14-
demethylase, which
participates in sterol synthesis. Sterols, (e.g., ergosterol in fungi) are
part of cell walls and necessary
for membrane structure and formation. Each triazole may vary in its action
within the sterol-
production pathway; however, the results are generally similar: abnormal
fungal growth and death
as a result of cell membrane deformities. Because the mode of action of
triazole is highly specific,
i.e., it targets only a single pathway in the fungus, there are instances
where mutations can occur in
certain fungal species that can make them resistant to triazoles, especially
in fungi that reproduce
rapidly such as rusts. If such a resistant strain occurs, repeated application
of the triazole can lead to
a buildup of a triazole -resistant subpopulation in an entire crop/plantation.
There are two types of
fungicide resistance: quantitative and qualitative. Quantitatively resistant
pathogens are less
sensitive to the fungicide compared to the wild type, but can still be
controlled with a higher use
rate and/or more frequent applications. On the other hand, qualitatively
resistant strains are
insensitive/unresponsive to the fungicide and can no longer be controlled at
labeled field rates. To
slow the rate of proliferation of resistant strains, it is useful to limit the
consecutive applications of
triazole fungicides to the earlier stages of fungal infection as well as
applying a second type of
fungicide that possesses another mode of action. It is therefore useful to
provide triazole
formulations that can easily be mixed with another type of fungicide (e.g., a
strobilurin) that has a
different mode of action to help reduce the risk of resistant strains. In
addition, improved
formulations that are more effective at lower rates, show longer-lasting
activity, or can be applied
less frequently due to improvements in systemic activity as well as decreasing
the potential for the
development of fungicide resistance.
Plant uptake and weak systemic effect
Fungicides can either be contact, translanninar or systemic. Contact
fungicides are not taken
up into the plant tissue, and only protect the plant where the spray is
deposited. Translaminar
fungicides redistribute the fungicide from the upper, sprayed leaf surface to
the lower, unsprayed
surface of the same leaf. Systemic fungicides are taken up and redistributed
through the xylem
vessels to the upper parts of the plant. Systemic activity is necessary to
provide curative
performance for a fungicide. Further, some triazoles are somewhat translaminar
(spreading through
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
14
individual leaves) and to a certain extent, weakly systemic (e.g., curative)
fungicides. Because of
these traits Triazoles are known to have primary curative activity, but are
disfavored in preventative
application.
When the triazole is applied to the plant, most of the active ingredient is
initially held on or
within the plant surface. If the triazole is showing weak systemic activity,
this is because the active
ingredient penetrates into the underlying plant cells (translaminar movement)
and also moves to
local zones above the point of uptake (local systemization via the xylem in
the leaf). The uptake of
the triazole into the cells of the leaf following application is dependent on
several factors: the
formulation type, active ingredient particle size, the additives/adjuvants
used in the formulation, the
other active ingredients mixed in or with the formulation, the target crop
(leaf type, surface,
weathering and plant age) and environmental factors that influence the drying
of the spray droplet.
Lack of, or low system effect can be problematic, as it means that any plant
tissue that needs
to be protected by the triazole formulation needs to be efficiently covered
during the application
process (typically spray). Unfortunately, aerial spray or foliar spray is
often non-uniform and does
not lead to complete coverage of the exterior of the plant (e.g., see Henriet
and Baur, Bayer
CropScience Journal 62(2):243, 2009). In addition, as plants grow they develop
new foliar tissue that
was not treated with the triazole and hence will not be protected from fungal
infection until the next
application. The degree of systemic activity can be demonstrated by evaluating
the performance of
the triazole for curative activity; improvements in curative activity can be
correlated with
improvements in systemization.
If a triazole could be made more systemic through improvements in formulation
it would
dramatically improve the impact of triazoles on target crops because of the
potentially reduced
application rates and enhanced efficacy (e.g., increase yields) of such
formulations.
Plant Health and Hidden Disease
Growers strive to obtain high yielding and high quality plants and crops.
Toward this goal,
agricultural strategies are utilized to maintain, optimize, and enhance plant
health from the time of
planting through to harvest. As a descriptive term, plant health refers to the
overall condition of a
plant, including its size, sturdiness, optimum maturity, consistency in growth
pattern and
reproductive activity. Growers often also define plant health in terms of
nneasureable outputs, such
as enhanced crop yield and economic return on production input.
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
As the effective control of fungal disease is of central importance in
improving and
optimizing plant health and crop yield, triazole fungicides are often applied
as part of regimes
directed towards achieving these results. Plant health applications of
triazoles may include curative
inoculations to control disease, inoculations for the purpose of combating
hidden disease,
inoculations under conditions that are favorable for the development of
disease (e.g., favorable
weather conditions), insurance applications, and other applications to improve
crop yield and
quality. Furthermore, environmental conditions are closely and constantly
monitored by growers,
and upon tending towards circumstances that are favorable for fungal
infections, triazole
applications are performed.
Of central importance to the improvement of plant health via the application
of triazole
fungicides is combating hidden or undiagnosed disease. Growers have implicated
hidden diseases
(i.e., cases in which the crop has below detection limit or non-obvious fungal
infection) in reduced
and variable crop yields. In response, triazole fungicides are often used in
plant health applications
such as insurance applications (e.g., applications that are made regardless of
disease pressure),
particularly on high potential crops frequently mixed with another fungicide
with a different mode of
action. In many cases these have been found to reverse or dampen the effects
of hidden disease on
crops and improve yield.
There are, however, persistent challenges related to the use of triazoles in
improving plant
health by combating hidden disease, the most problematic of which are related
to correct timing of
application and low or insufficient levels of curative activity. For example,
prior to early triazole
applications (e.g., the first application of the season), there is often a
level of latent infection or
hidden disease in the crop. In such cases, commercial formulations that
demonstrate preventative
activity but that suffer from low or less than adequate levels of curative
activity would be ineffective
at improving plant health by combating hidden disease and even fungicides with
curative properties
could be made more efficient. To compensate in part for their low or
inefficient curative activity,
commercial formulations are sometimes applied at increased rates. Furthermore,
plant physiology
and pathology are extremely complex, and there remain unanswered questions
surrounding the
optimal time points for application of fungicides to improve plant health and
risks of fungicide
resistance by combating hidden disease.
Related to the complexity of plant physiology and influencing plant health is
the fact that
triazoles can function as plant growth regulators. Briefly, plant growth
regulators are man-made
chemical compounds that effect the growth and development of plants in some
way. Naturally
synthesized compound, either from the plant itself or from another source
within the plant's
environment (e.g., bacteria) are typically called plant hormones. Plant growth
regulators can
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
16
manifest themselves in a wide variety of ways within a plant as the plant
grows. Some of the effects
can be beneficial or detrimental to the plant from a plant health perspective
and the same triazole
compound may produce a mix of beneficial and detrimental effects in a given
plant. For example,
some plant growth regulators reduce the size and weight of stems and leaves of
a plant. Some other
plant growth regulators produce higher cell density in a plant's leaves, or
increased resistance to
stress conditions (e.g., drought, chilling). The specific results and effects
of a plant growth regulator
depend on many factors including the particular regulator, the particular
plant, the environmental
conditions and the time of application.
Triazoles are known to act as plant growth regulators, in addition to their
fungicidal uses.
Various plant growth effects from triazoles have been described including
increased cell density,
increased chlorophyll density, increased leaf thickness and vibrancy, among
other effects. Some
triazoles have been shown to stunt the growth of some plants either stem and
leaf length or weight.
Primarily triazoles as plant growth regulators disrupt the gibberellin
pathways. Because triazoles
provide the additional benefits beyond fungicide applications they can have a
more pronounced
effect on overall plant health, as shown by increased yields. Triazoles' role
as plant growth
regulators can help combat hidden disease, stunt the growth of pest/competing
plants, and trigger
various biological effects within the plant to improve overall plant health in
a variety of growth
conditions. Improved triazole formulation can lead to enhanced plant growth
regulator effects as
well. Triazole formulations with improved water solubility, improved systemic
effect or greater
residual activity can have great regulator effects, leading to improved plant
health. Improved plant
health, in turn, can lead to higher product yields.
It would thus be desirable to develop triazole formulations that provide
increased levels of
curative activity for plant health applications, including the treatment of
latent and hidden fungal
disease. For example, it would be useful to produce triazole formulations that
have increased levels
of curative activity by imparting greater systemic properties to a triazole or
improving the systemic
properties of the fungicide. Such formulations would be more effective in
plant health applications
and could therefore be used at lower effective dose rates than currently
available commercial
formulations. Furthermore, it would be useful to provide triazole formulations
that could in part
mitigate the difficulties associated with correct timing of fungicide
applications directed to
improving plant health. For example, formulations that display enhanced
residual activity would
increase the window of opportunity for successful application timing. Lastly,
it would be useful to
provide triazole formulations that could improve plant health by having a
plant growth regulator
effect. Plant yields can be further improved by providing a formulation that
could provide a number
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
17
of the functions described above (e.g., improved translaminar activity,
improved plant growth
regulator effect, improved residual activity).
Formulations ¨ Generally
Several synthetic triazoles (including difenoconazole, fenbuconazole,
myclobutanil,
propiconazole, tebuconazole, tetraconazole, triticonazole and epiconazole)
formulations are now
available commercially, the bulk of which are used in agricultural
applications. Despite a common
mode of action, triazoles exhibit definite practical differences, e.g.,
different mobility in the plant.
The aforementioned limitations of triazoles, and their formulations, when used
as fungicides
manifest themselves in (a) how they are currently applied to plants and (b)
how they are formulated
by manufacturers. As an example, because triazoles are susceptible to
degradation (either from
photolysis or exposure of field conditions) end users (e.g., farmers or golf
course maintenance
managers) need to apply triazoles more often than if they were longer lasting.
As another example,
because some triazoles lack systemic activity, or have limited system activity
(which would help
protect new growth of crops), end users need to continually re-apply triazoles
in order to protect
crops from fungal infection. Furthermore, because of the inherent threat of
forming triazole
resistant strains, end users need triazole formulations that that can be
easily mixed with other types
of formulated fungicides as well as formulations that have improved residual
activity (i.e., would
need less applications). These limitations are compounded by increasing
pressure on end users who
are faced with increasing regulatory and consumer pressure to use fewer
pesticides and/or
fungicides and in lower quantities.
In order to address these limitations, a variety of complicated formulation
techniques and
formulation agents have been developed to counter to the UV instability, water
insolubility, non-
systemic nature, and other limitations of triazoles.
In order for a triazole to be efficiently applied to a plant or fungus, the
triazole product
needs to be dispersible in water. Two common formulation techniques to do this
are to produce
either an emulsifiable concentrate (EC) or a suspension concentrate (SC). An
EC is a formulation
where the active ingredient is dissolved in a suitable solvent in the presence
of surfactants. When
the EC is dispersed into the spray tank and agitated, the surfactants emulsify
the solvent into water,
and the active ingredient is delivered in the solvent phase to the plant or
fungus. ECs frequently do
not require, or are incompatible with, the addition of surfactant in the spray
tank. Because ECs
contain solvent and significant amounts of surfactant in the formulation,
additional surfactant
increases the formulations' phytotoxicity. Even without the increased danger
to the plant itself, the
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
18
formulation would not like exhibit an improvement in agrochemical performance.
A SC is a high-solids concentrate in water. The active ingredient is milled
into particles that
are 1-10 microns (Alan Knowles, Agrow Reports: New Developments in Crop
Protection Product
Formulation. London: Agrow Reports May 2005). These solid particles are then
dispersed into water
at high concentration using surfactants. After adding the SC into the spray
tank, the surfactant-
stabilized particles disperse into water and are applied (still as solid
particles) to the leaf surface.
Other common formulation techniques used for some crop protection active
ingredients include
nnicroencapsulations (CS) and emulsions (EW or OW). Solid formulation
techniques that are
currently used include water-dispersible granules (WG) or powders (WP), where
the active
ingredient is absorbed to a dispersible carrier that is provided dry to the
farmer. When mixed into
the spray tank, the carrier disperses into the water, carrying the active
ingredient with it. Particle
sizes for these carriers can be anywhere in the range of 1-10 microns (Alan
Knowles, Agrow Reports:
New Developments in Crop Protection Product Formulation. London: Agrow Reports
May 2005).
As an alternative to these approaches, we have developed new classes triazole
formulations.
As demonstrated in the Examples and as discussed below, in some embodiments
these new triazole
formulations are more dispersible in water and have enhanced stability (i.e.,
longer lasting). In some
embodiments, these new triazole formulations have increased curative
(systemic) and preventative
performance as compared to existing formulations. Further, the new
formulations are also
compatible with other agricultural products (surfactants, leaf wetters,
fertilizers, etc.), and are stable
in non-ideal solution conditions such high salt, extreme pH, hard water,
elevated temperatures, etc.
These enhancements/improvements in the formulation can also help address the
resistance of some
fungi by being (1) compatible with a second fungicide, either tank-mixed or
pre-mixed in the original
formulation and (2) requiring less fungicide in each application as well as
improved efficacy and
reduced application rates. In general, these new triazole formulations
comprise nanoparticles
(optionally in aggregate form) of polymer-associated triazoles along with
various formulating agents.
Additionally, because the instant formulations are based around nanoparticles
of polymer-
associated active ingredients, they are stable to relatively high salt
conditions. Stability in high salt
conditions is required especially when the formulation is to be mixed with
other secondary
agricultural products such as a concentrated fertilizer mix, exposed to high
salt conditions (e.g., used
in or with hard waters) mixed with other formulations (other pesticides,
fungicides, and herbicides)
or mixed with other tank-mix adjuvants. The ability to mix our formulations
with other products can
be beneficial to the end user because simultaneous agricultural products can
be applied in a single
application.
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
19
Formulations ¨ Components
In various aspects, the present disclosure provides formulations that comprise
nanoparticles
(optionally in aggregate form) of polymer-associated active ingredient along
with various
formulating agents.
Active Ingredient
As used herein, the term "active ingredient" ("al", "Al", "al.", "Al.") refers
to triazole
compounds (i.e., triazoles). Structurally, the basic common feature in this
family is the presence of
triazole heterocyclic structure. Many triazoles include a triazole group:
Often, though not always, in conjunction with a halogen substituted phenyl
group. For
example, difenoconazole, which structure is shown below, includes both groups.
CH3
0
0
CI
c"NN
Non-limiting examples of triazole fungicides include azaconazole (1-([2-(2,4-
dichloropheny1)-
1,3-dioxolan-2-yl]methyll-1H-1,2,4-triazole), Bronnuconazole (1-
[(2R5,4RS;2R5,45R)-4-bronno-2-(2,4-
dichlorophenyl)tetrahydrofurfury1]-1H-1,2,4-triazole), cyproconazole
((2RS,3RS;2RS,3SR)-2-(4-
chloropheny1)-3-cyclopropy1-1-(1H-1,2,4-triazol-1-yl)butan-2-ol),
diclobutrazol ((2R5,3R5)-1-(2,4-
dichloropheny1)-4,4-dinnethyl-2-(1H-1,2,4-triazol-1-yl)pentan-3-ol),
difenoconazole (3-chloro-4-
[(2R5,4R.S;2RS,4SR)-4-methyl-2-(1H-1,2,4-triazol-1-ylmethyl)-1,3-dioxolan-2-
yl]phenyl 4-chlorophenyl
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
ether), diniconazole ((E)-(RS)-1-(2,4-dichloropheny1)-4,4-dimethyl-2-(1H-1,2,4-
triazol-1-yppent-1-en-
3-ol), epoxiconazole ((2RS,3SR)-1-[3-(2-chlorophenyI)-2,3-epoxy 2 (4
fluorophenyl)propyI]-1H-1,2,4-
triazole), etaconazole (1-[(2RS,4RS;2RS,4SR)-2-(2,4-dichloropheny1)-4-ethyl-
1,3-dioxolan-2-ylmethyl]-
1H-1,2,4-triazole), fenbuconazole ((RS) 4 (4 chloropheny1)-2-pheny1-2-(1H-
1,2,4-triazol-1-
ylmethyl)butyronitrile), fluquinconazole (3-(2,4-dichlorophenyI)-6-fluoro-2-
(1H-1,2,4-triazol-1-
yl)quinazolin-4(3H)-one), flusilazole (bis(4-fluorophenyl)(nnethyl)(1H-1,2,4-
triazol-1-ylmethyl)silane
or 1-{[bis(4-fluorophenyl)(methypsilyl]methy11-1H-1,2,4-triazole), flutriafol
((RS)-2,4'-difluoro-a-(1H-
1,2,4-triazol-1-yInnethyl)benzhydryl alcohol), furconazole ((2RS,5RS;2RS,5SR)-
5-(2,4-
dichlorophenyptetrahydro-5-(1H-1,2,4-triazol-1-ylmethyl)-2-furyl 2,2,2-
trifluoroethyl ether),
hexaconazole ((RS)-2-(2,4-dichlorophenyI)-1-(1H-1,2,4-triazol-1-yl)hexan-2-
ol), innibenconazole (4-
chlorobenzyl (EZ)-N-(2,4-dichlorophenyI)-2-(1H-1,2,4-triazol-1-
yl)thioacetamidate), ipconazole
((1RS,2SR,5RS;1RS,2SR,5SR)-2-(4-chlorobenzy1)-5-isopropy1-1-(1H-1,2,4-triazol-
1-
ylmethyl)cyclopentanol), metconazole U1RS,5RS;1RS,5SR)-5-(4-chlorobenzy1)-2,2-
dimethyl-1-(1H-
1,2,4-triazol-1-ylnnethyl)cyclopentanol), myclobutanil ((RS)-2-(4-
chlorophenyI)-2-(1H-1,2,4-triazol-1-
ylmethyl)hexanenitrile), penconazole ((RS)-142-(2,4-dichlorophenyppenty1]-1H-
1,2,4-triazole),
propiconazole ((2RS,4RS;2RS,4SR)-142-(2,4-dichloropheny1)-4-propy1-1,3-
dioxolan-2-ylmethyl]-1H-
1,2,4-triazole), prothioconazole URS)-2-[2-(1-chlorocyclopropy1)-3-(2-
chloropheny1)-2-
hydroxypropyl]-2,4-dihydro-1,2,4-triazole-3-thione), quinconazole (3-(2,4-
dichlorophenyI)-2-(1H-
1,2,4-triazol-1-y1)-quinazolin-4(3H)-one), sinneconazole URS)-2-(4-
fluoropheny1)-1-(1H-1,2,4-triazol-1-
y1)-3-(trinnethylsilyl)propan-2-01), tebuconazole ((RS)-1-p-chloropheny1-4,4-
dinnethy1-3-(1H-1,2,4-
triazol-1-yInnethyl)pentan-3-01), tetraconazole ((RS)-2-(2,4-dichlorophenyI)-3-
(1H-1,2,4-triazol-1-
yl)propyl 1,1,2,2-tetrafluoroethyl ether), triadimenfon ((RS)-1-(4-
chlorophenoxy)-3,3-dinnethy1-1-(1H-
1,2,4-triazol-1-yl)butan-2-one), triadinnenol ((1RS,2RS;1RS,2SR)-1-(4-
chlorophenoxy)-3,3-dimethyl-1-
(1H-1,2,4-triazol-1-yl)butan-2-ol), triticonazole URS)-(E)-5-(4-
chlorobenzylidene)-2,2-dimethy1-1-(1H-
1,2,4-triazol-1-ylnnethyl)cyclopentanol), uniconazole ((E)-(RS)-1-(4-
chloropheny1)-4,4-dinnethy1-2-(1H-
1,2,4-triazol-1-yl)pent-1-en-3-o1).
In some embodiments, triazole formulations are applied in combination with one
or more
other pesticides (e.g., insecticides, herbicides, fungicides). For example,
the triazole formulations
can be applied with other fungicides with a different mode of action as
compared to the triazole
(e.g., strobilurin). Such mixed applications are typically used to mitigate
the potential development
of fungicide resistance to a particular fungicide in the targeted fungi.
Exemplary strobilurins include,
but are not limited to, azoxystrobin, picoxystrobin, pyraclostrobin,
orysastrobin, nnetonninostrobin
and trifloxystrobin. The second fungicide may be a completely separate
formulation, mixed with a
triazole formulation by the grower in the application tank. In some
embodiments, the triazole and
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
21
second fungicide (e.g., a triazole) are mixed together in a single
formulation, which is applied (or
diluted and applied) by a user.
For example, the additional pesticide (e.g., fungicide) can make up between
about 0.5 and
about 20 weight %, about 0.5 and about 10 weight %, between about 0.5 and
about 5 weight %,
between about 0.5 and about 3 weight %, between about 1 and about 30 weight %,
between about
1 and about 20 weight %, between about 1 and about 10 weight %, between about
1 and about 5
weight %, between about 2 and about 30 weight %, between about 2 and about 20
weight %,
between about 2 and about 10 weight %, between about 2 and about 5 weight %,
between about 3
and about 30 weight %, between about 3 and about 20 weight %, between about 3
and about 10
weight %, between about 3 and about 5 weight %, between about 5 and about 30
weight %,
between about 5 and about 20 weight %, between about 5 and about 10 weight %
of the
formulation. In some embodiments, the additional pesticide (e.g., fungicide)
can make up between
about 0.1 and 1 weight % of the formulation, between about 0.1 and 2 weight %
of the formulation
between about 0.1 and 3 weight % of the formulation between about 0.1 and 5
weight % of the
formulation, between about 0.1 and 10 weight % of the formulation.
Nanoparticles of polymer-associated active ingredient
As used herein, the terms "nanoparticles of polymer-associated active
ingredient",
"nanoparticles of polymer-associated triazole compound" or "active ingredient
associated with polymer
nanoparticles" refer to nanoparticles comprising one or more collapsed
polymers that are associated
with the active ingredient. In some embodiments the collapsed polymers are
cross-linked. As
discussed below, in some embodiments, our formulations may include aggregates
of nanoparticles.
Exemplary polymers and methods of preparing nanoparticles of polymer-
associated active
ingredient are described more fully below.
In some embodiments, the active ingredient is associated with preformed
polymer
nanoparticles. The associating step may involve dispersing the polymer
nanoparticles in a first
solvent and then dispersing the active ingredient in a second solvent that is
miscible or partially
miscible with the first solvent, mixing the two dispersions and then either
removing the second or
first solvent from the final mixture. In some embodiments, all the solvent is
removed by vacuum
evaporation, freeze drying or spray drying. The associating step may also
involve dispersing both the
preformed polymer nanoparticles and active ingredients in a common solvent and
removing all or a
portion of the common solvent from the final mixture.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
22
In some embodiments, the associating step may involve milling the active
ingredient in the
presence of pre-formed polymer nanoparticles. It is surprising that if the
active ingredient alone is
milled under these conditions; the resulting particle size is significantly
larger than if it is milled in
the presence of pre-formed polymer nanoparticles. In general, size reduction
processes such as
milling do not enable the production of particle sizes that are produced via
milling in the presence of
nanoparticles of the current disclosure. Without wishing to be bound by any
theory, it is thought
that interaction between the active ingredient and the nanoparticles during
the milling process
facilitates the production of smaller particles than would be formed via
milling in the absence of the
nanoparticles.
Non-limiting examples of milling methods that may be used for the association
step can be
found in U.S. Patent No. 6,6046,98 and include ball milling, bead milling, jet
milling, media milling,
and homogenization, as well as other milling methods known to those of skill
in the art. Non-limiting
examples of mills that can be for the association step include attritor mills,
ball mills, colloid mills,
high pressure homogenizers, horizontal mills, jet mills, swinging mills, and
vibratory mills. In some
embodiments, the associating step may involve milling the active ingredient in
the presence of pre-
formed polymer nanoparticles and an aqueous phase. In some embodiments, the
associating step
may involve wet or dry milling of the active ingredient in the presence of pre-
formed nanoparticles.
In some embodiments, the association step may involve milling the active
ingredient and pre-formed
polymer nanoparticles in the presence of one or more formulating agents.
In general and without limitation, the active ingredient may be associated
with regions of
the polymer nanoparticle that elicit a chemical or physical interaction with
the active ingredient.
Chemical interactions can include hydrophobic interactions, affinity pair
interactions, H-bonding, and
van der Waals forces. Physical interactions can include entanglement in
polymer chains and/or
inclusion within the polymer nanoparticle structure. In some embodiments, the
active ingredient
can be associated in the interior of the polymer nanoparticle, on the surface
of the polymer
nanoparticle, or both the surface and the interior of the polymer
nanoparticle. Furthermore, the
type of association interactions between the active ingredient and the polymer
nanoparticle can be
probed using spectroscopic techniques such as NMR, IR, UV-vis, and emission
spectroscopies. For
example, in cases where the triazole active ingredient is normally crystalline
when not associated
with the polymer nanoparticles, the nanoparticles of polymer-associated
triazole compounds
typically do not show the endothermic melting peak or show a reduced
endothermic melting peak of
the pure crystalline active ingredient as seen in differential thermal
analysis (DTA) or differential
scanning calorimetry (DSC) measurements
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
23
Nanoparticles of polymer-associated active ingredients can be prepared with a
range of
average diameters, e.g., between about 1 nm and about SOO nm. The size of the
nanoparticles can
be adjusted in part by varying the size and number of polymers that are
included in the
nanoparticles. In some embodiments, the average diameter ranges from about 1
nm to about 10
nm, from about 1 nm to about 20 nm, from about 1 nm to about 30 nm, from about
1 nm to about
50 nm, from about 10 nm to about 50 nm, from about 10 nm to about 100 nm, from
about 20 nm to
about 100 nm, from about 20 nm to about 100 nm, from about 50 nm to about 200
nm, from about
50 nm to about 250 nm, from about 50 nm to about 300 nm, from about 100 nm to
about 250 nm,
from about 100 nm to about 300 nm, from about 200 nm to about 300 nm, from
about 200 nm to
about 500 nm, from about 250 nm to about 500 nm, and from about 300 nm to
about 500 nm.
These and other average diameters described herein are based on volume average
particle sizes that
were measured in solution by dynamic light scattering on a Malvern Zetasizer
ZS in CIPAC D water,
0.1M NaCI, or in deionized water at 200 ppm active concentration. Various
forms of microscopies
can also be used to visualize the sizes of the nanoparticles such as atomic
force microscopy (AFM),
transmission electron microscopy (TEM), scanning electron microscopy (SEM) and
optical
microscopy.
In some embodiments, the aggregates of nanoparticles of polymer-associated
active
ingredients have an average particle size between about 10 nm and about 5,000
nm when dispersed
in water under suitable conditions. In some embodiments, the aggregates have
an average particle
size between about 10 nm and about 1,000 nm. In some embodiments, the
aggregates have an
average particle size between about 10 nm and about 500 nm. In some
embodiments, the
aggregates have an average particle size between about 10 nm and about 300 nm.
In some
embodiments, the aggregates have an average particle size between about 10 nm
and about 200
nm. In some embodiments, the aggregates have an average particle size between
about 50 nm and
about 5,000 nm. In some embodiments, the aggregates have an average particle
size between about
50 nm and about 1,000 nm. In some embodiments, the aggregates have an average
particle size
between about 50 nm and about 500 nm. In some embodiments, the aggregates have
an average
particle size between about 50 nm and about 300 nm. In some embodiments, the
aggregates have
an average particle size between about 50 nm and about 200 nm. In some
embodiments, the
aggregates have an average particle size between about 100 nm and about 5,000
nm. In some
embodiments, the aggregates have an average particle size between about 100 nm
and about 1,000
nm. In some embodiments, the aggregates have an average particle size between
about 100 nm and
about 500 nm. In some embodiments, the aggregates have an average particle
size between about
100 nm and about 300 nm. In some embodiments, the aggregates have an average
particle size
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
24
between about 100 nm and about 200 nm. In some embodiments, the aggregates
have an average
particle size between about SOO nm and about 5000 nm. In some embodiments, the
aggregates have
an average particle size between about SOO nm and about 1000 nm. In some
embodiments, the
aggregates have an average particle size between about 1000 nm and about 5000
nm. Particle size
can be measured by the techniques described above.
As described in detail in the examples, in some embodiments, pre-formed
polymer
nanoparticles that have been associated with active ingredient to generate
nanoparticles or
aggregates of nanoparticles of polymer-associated active ingredients
(associated nanoparticles) can
be recovered after extraction of the active ingredient. In some embodiments,
the active ingredient
can be extracted from nanoparticles or aggregates of nanoparticles of polymer-
associated active
ingredient by dispersing the associated nanoparticles in a solvent that
dissolves the active ingredient
but that is known to disperse the un-associated, preformed nanoparticles
poorly or not at all. In
some embodiments, after extraction and separation, the insoluble nanoparticles
that are recovered
have a size that is smaller than the nanoparticles or aggregates of
nanoparticles of polymer-
associated active ingredients as measured by DLS. In some embodiments, after
extraction and
separation, the insoluble nanoparticles that are recovered have a size that is
similar or substantially
the same as the size of original pre-formed polymer nanoparticles (prior to
association) as measured
by DLS. In some embodiments, the nanoparticles are prepared from
poly(nnethacrylic acid-co-ethyl
acrylate). In some embodiments, the active ingredient is difenoconazole. In
some embodiments, the
extraction solvent is acetonitrile.
It should be understood that the association step to generate nanoparticles of
polymer
associated active ingredient need not necessarily lead to association of the
entire fraction the active
ingredient in the sample with pre-formed polymer nanoparticles (not all
molecules of the active
ingredient in the sample must be associated with polymer nanoparticles after
the association step).
Likewise, the association step need not necessarily lead to the association of
the entire fraction of
the pre-formed nanoparticles in the sample with active ingredient (not all
nanoparticle molecules in
the sample must be associated with the active ingredient after the association
step).
Similarly, in formulations comprising nanoparticles of polymer-associated
active, the entire
fraction of active ingredient in the formulation need not be associated with
pre-formed polymer
nanoparticles (not all molecules of the active ingredient in the sample must
be associated with
polymer nanoparticles in the formulation). Likewise, in formulations
comprising nanoparticles of
polymer-associated active ingredient, the entire fraction of pre-formed
polymer nanoparticles in the
formulation need not be associated with active ingredient (not all of
nanoparticle molecules in the
sample must be associated with the active ingredient in the formulation).
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
In some embodiments, the nanoparticles are prepared using a polymer that is a
polyelectrolyte. Polyelectrolytes are polymers that contain monomer units of
ionized or ionizable
functional groups. They can be linear, branched, hyperbranched or dendrimeric,
and they can be
synthetic or naturally occurring. Ionizable functional groups are functional
groups that can be
rendered charged by adjusting solution conditions, while ionized functional
group refers to chemical
functional groups that are charged regardless of solution conditions. The
ionized or ionizable
functional group can be cationic or anionic, and can be continuous along the
entire polymer chain
(e.g., in a honnopolymer), or can have different functional groups dispersed
along the polymer chain,
as in the case of a co-polymer (e.g., a random co-polymer). In some
embodiments, the polymer can
be made up of monomer units that contain functional groups that are either
anionic, cationic, both
anionic and cationic, and can also include other monomer units that impart a
specific desirable
property to the polymer.
In some embodiments, the polyelectrolyte is a honnopolymer. Non limiting
examples of
honnopolynner polyelectrolytes include: poly(acrylic acid), poly(methacrylic
acid), poly(styrene
sulfonate), poly(ethyleneinnine), chitosan, poly(dimethylannnnoniunn
chloride), poly(allylannine
hydrochloride), and carboxymethyl cellulose.
In some embodiments, the polyelectrolyte is a co-polymer. Non limiting
examples of co-
polymer polyelectrolytes include: poly(methacrylic acid-co-ethyl acrylate);
poly(methacrylic acid-co-
styrene); poly(methacrylic acid-co-butylnnethacrylate); poly[acrylic acid-co-
poly(ethylene glycol)
methyl ether methacrylate].
In some embodiments, the polyelectrolyte can be made from one or more monomer
units to
form homopolynners, copolymers or graft copolymers of: ethylene; ethylene
glycol; ethylene oxide;
carboxylic acids including acrylic acid, methacrylic acid, itaconic acid, and
nnaleic acid;
polyoxyethylenes or polyethyleneoxide; and unsaturated ethylenic mono or
dicarboxylic acids; lactic
acids; amino acids; amines including dinnethlyannnnoniunn chloride,
allylannine hydrochloride;
nnethacrylic acid; ethyleneimine; acrylates including methyl acrylate, ethyl
acrylate, propyl acrylate,
n-butyl acrylate ("BA"), isobutyl acrylate, 2-ethyl acrylate, and t-butyl
acrylate; nnethacrylates
including ethyl nnethacrylate, n-butyl nnethacrylate, and isobutyl
nnethacrylate; acrylonitriles;
nnethacrylonitrile; vinyls including vinyl acetate, vinylversatate,
vinylpropionate, vinylfornnannide,
vinylacetannide, vinylpyridines, and vinyllinnidazole; vinylnapthalene,
vinylnaphthalene sulfonate,
vinylpyrrolidone, vinyl alcohol; anninoalkyls including anninoalkylacrylates,
aminoalkylsnnethacrylates,
and anninoalkyl(meth)acrylamides; styrenes including styrene sulfonate; d-
glucosannine; glucaronic
acid-N-acetylglucosannine; N-isopropylacrylannide; vinyl amine. In some
embodiments, the
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
26
polyelectrolyte polymer can include groups derived from polysaccharides such
as dextran, gums,
cellulose, or carboxynnethyl cellulose.
In some embodiments, the polyelectrolyte comprises poly(methacrylic acid-co-
ethyl
acrylate) polymer. In some embodiments, the mass ratio of nnethacrylic acid to
ethyl acrylate in the
poly(methacrylic acid-co-ethyl acrylate) polymer is between about 50:50 and
about 95:5. In some
embodiments, the mass ratio of nnethacrylic acid to ethyl acrylate in the
poly(methacrylic acid-co-
ethyl acrylate) polymer is between about 70:30 and about 95:5. In some
embodiments, the mass
ratio of methacrylic acid to ethyl acrylate in the poly(methacrylic acid-co-
ethyl acrylate) polymer is
between about 80:20 and about 95:5. In some embodiments, the mass ratio of
nnethacrylic acid to
ethyl acrylate in the poly(methacrylic acid-co-ethyl acrylate) polymer is
between about 85:15 and
about 95:5. In some embodiments, the mass ratio of nnethacrylic acid to ethyl
acrylate in the
poly(methacrylic acid-co-ethyl acrylate) polymer is between about 60:40 and
about 80:20.
In some embodiments, the polyelectrolyte comprises poly(methacrylic acid-co-
styrene)
polymer. In some embodiments, the mass ratio of methacrylic acid to styrene in
the
poly(methacrylic acid-co-styrene) polymer is between about 50:50 and about
95:5. In some
embodiments, the mass ratio of nnethacrylic acid to styrene in the
poly(methacrylic acid-co-styrene)
polymer is between about 70:30 and about 95:5. In some embodiments, the mass
ratio of
nnethacrylic acid to styrene in the poly(methacrylic acid-co-styrene) polymer
is between about 80:20
and about 95:5. In some embodiments, the mass ratio of nnethacrylic acid to
styrene in the
poly(methacrylic acid-co-styrene) polymer is between about 85:15 and about
95:5. In some
embodiments, the mass ratio of nnethacrylic acid to styrene in the
poly(methacrylic acid-co-styrene)
polymer is between about 60:40 and about 80:20.
In some embodiments, the mass ratio of methacrylic acid to butyl methacrylate
in the
poly(methacrylic acid-co-butylnnethacrylate) polymer is between about 50:50
and about 95:5. In
some embodiments, the mass ratio of nnethacrylic acid to butyl nnethacrylate
in the poly(methacrylic
acid-co-butylmethacrylate) polymer is between about 70:30 and about 95:5. In
some embodiments,
the mass ratio of nnethacrylic acid to butyl nnethacrylate in the
poly(methacrylic acid-co-
butylnnethacrylate) polymer is between about 80:20 and about 95:5. In some
embodiments, the
mass ratio of nnethacrylic acid to butyl nnethacrylate in the poly(methacrylic
acid-co-
butylnnethacrylate) polymer is between about 85:15 and about 95:5. In some
embodiments, the
mass ratio of nnethacrylic acid to butyl nnethacrylate in the poly(methacrylic
acid-co-
butylnnethacrylate) polymer is between about 60:40 and about 80:20.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
27
In some embodiments, the homo or co-polymer is water soluble at pH 7. In some
embodiments, the polymer has solubility in water above about 1 weight %. In
some embodiments,
the polymer has solubility in water above about 2 weight %. In some
embodiments, the polymer has
solubility in water above about 3 weight %. In some embodiments, the polymer
has solubility in
water above about 4 weight %. In some embodiments, the polymer has solubility
in water above
about 5 weight %. In some embodiments, the polymer has solubility in water
above about 10 weight
%. In some embodiments, the polymer has solubility in water above about 20
weight %. In some
embodiments, the polymer has solubility in water above about 30 weight %. In
some embodiments,
the polymer has solubility in water between about 1 and about 30 weight %. In
some embodiments,
the polymer has solubility in water between about 1 and about 10 weight %. In
some embodiments,
the polymer has solubility in water between about 5 and about 10 weight %. In
some embodiments,
the polymer has solubility in water between about 10 and about 30 weight %. In
some
embodiments the solubility of the polymer in water can also be adjusted by
adjusting pH or other
solution conditions in water.
In some embodiments, the polyelectrolyte polymer has a weight average (Mw)
molecular
weight between about 5,000 and about 4,000,000 Daltons. In some embodiments,
the
polyelectrolyte polymer has a weight average molecular weight between about
100,000 and about
2,000,000 Daltons. In some embodiments, the polyelectrolyte polymer has a
weight average
molecular weight between about 100,000 and about 1,000,000 Daltons. In some
embodiments, the
polyelectrolyte polymer has a weight average molecular weight between about
100,000 and about
750,000 Daltons. In some embodiments, the polyelectrolyte polymer has a weight
average
molecular weight between about 100,000 and about 500,000 Daltons. In some
embodiments, the
polyelectrolyte polymer has a weight average molecular weight between about
100,000 and about
200,000 Daltons. In some embodiments, the polyelectrolyte polymer has a weight
average
molecular weight between about 200,000 and about 2,000,000 Daltons. In some
embodiments, the
polyelectrolyte polymer has a weight average molecular weight between about
200,000 and about
1,000,000 Daltons. In some embodiments, the polyelectrolyte polymer has a
weight average
molecular weight between about 200,000 and about 500,000 Daltons. In some
embodiments, the
polyelectrolyte polymer has a weight average molecular weight between about
300,000 and about
2,000,000 Daltons. In some embodiments, the polyelectrolyte polymer has a
weight average
molecular weight between about 300,000 and about 1,000,000 Daltons. In some
embodiments, the
polyelectrolyte polymer has a weight average molecular weight between about
300,000 and about
500,000 Daltons. In some embodiments, the polyelectrolyte polymer has a weight
average molecular
weight between about 5,000 and about 250,000 Daltons. In some embodiments, the
polyelectrolyte
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
28
polymer has a weight average molecular weight between about 5,000 and about
50,000 Daltons. In
some embodiments, the polyelectrolyte polymer has a weight average molecular
weight between
about 5,000 and about 100,000 Daltons. In some embodiments, the
polyelectrolyte polymer has a
weight average molecular weight between about 5,000 and about 250,000 Daltons.
In some
embodiments, the polyelectrolyte polymer has a weight average molecular weight
between about
50,000 and about 250,000 Daltons.
In some embodiments, the apparent molecular weight of the polyelectrolyte
polymer (e.g.,
the molecular weight determined via certain analytical measurements such as
size exclusion
chromatography or DLS) is lower than the actual molecular weight of a polymer
due to crosslinking
within the polymer. In some embodiments, a crosslinked polyelectrolyte polymer
of the present
disclosure might have a higher actual molecular weight than the experimentally
determined
apparent molecular weight. In some embodiments, a crosslinked polyelectrolyte
polymer of the
present disclosure might be a high molecular weight polymer despite having a
low apparent
molecular weight.
Nanoparticles of polymer-associated active ingredients and/or aggregates of
these
nanoparticles can be part of a formulation in different amounts. The final
amount will depend on
many factors including the type of formulation (e.g., liquid or solid, granule
or powder, concentrated
or not, etc.). In some instances the nanoparticles (including both the polymer
and active ingredient
components) make up between about 1 and about 98 weight % of the total
formulation. In some
embodiments, the nanoparticles make up between about 1 and about 90 weight %
of the total
formulation. In some embodiments, the nanoparticles make up between about 1
and about 75
weight % of the total formulation. In some embodiments, the nanoparticles make
up between about
1 and about 50 weight % of the total formulation. In some embodiments, the
nanoparticles make up
between about 1 and about 30 weight % of the total formulation. In some
embodiments, the
nanoparticles make up between about 1 and about 25 weight % of the total
formulation. In some
embodiments, the nanoparticles make up between about 1 and about 10 weight %
of the total
formulation. In some embodiments, the nanoparticles make up between about 5
and about 15
weight % of the total formulation. In some embodiments, the nanoparticles make
up between about
and about 25 weight % of the total formulation. In some embodiments, the
nanoparticles make up
between about 10 and about 25 weight % of the total formulation. In some
embodiments, the
nanoparticles make up between about 10 and about 30 weight % of the total
formulation. In some
embodiments, the nanoparticles make up between about 10 and about 50 weight %
of the total
formulation. In some embodiments, the nanoparticles make up between about 10
and about 75
weight % of the total formulation. In some embodiments, the nanoparticles make
up between about
29
and about 90 weight % of the total formulation. In some embodiments, the
nanoparticles make
up between about 10 and about 98 weight % of the total formulation. In some
embodiments, the
nanoparticles make up between about 25 and about 50 weight % of the total
formulation. In some
embodiments, the nanoparticles make up between about 25 and about 75 weight %
of the total
formulation. In some embodiments, the nanoparticles make up between about 25
and about 90
weight % of the total formulation. In some embodiments, the nanoparticles make
up between about
30 and about 98 weight % of the total formulation. In some embodiments, the
nanoparticles make
up between about 50 and about 90 weight % of the total formulation. In some
embodiments, the
nanoparticles make up between about 50 and about 98 weight % of the total
formulation. In some
embodiments, the nanoparticles make up between about 75 and about 90 weight %
of the total
formulation. In some embodiments, the nanoparticles make up between about 75
and about 98
weight % of the total formulation.
In some embodiments, the nanoparticles of polymer-associated active
ingredients are
prepared according to a method disclosed in United States Patent Application
Publication No.
20100210465. In some
embodiments, polymer nanoparticles without active ingredients are made by
collapse of a
polyelectrolyte with a collapsing agent and then rendering the collapsed
conformation permanent
by intra-particle cross-linking. The active ingredient is then associated with
this pre-formed polymer
nanoparticle. In some embodiments, the formulation contains the same amount
(by weight) of
active ingredient and polymer nanoparticle, while in other embodiments the
ratio of active
ingredient to polymer nanoparticle (by weight) can be between about 1:10 and
about 10:1, between
about 1:10 and about 1:5, between about 1:5 and about 1:4, between about 1:4
and about 1:3,
between about 1:3 and about 1:2, between about 1:2 and about 1:1, between
about 1:5 and about
1:1, between about 5:1 and about 1:1, between about 2:1 and about 1:1, between
about 3:1 and
about 2:1, between about 4:1 and about 3:1, between about 5:1 and about 4:1,
between about 10:1
and about 5:1, between about 1:3 and about 3:1, between about 5:1 and about
1:1, between about
1:5 and about 5:1, or between about 1:2 and about 2:1.
As noted above, in some embodiments, the associating step may involve
dispersing the
polymer nanoparticles in a first solvent, dispersing the active ingredient in
a second solvent that is
miscible or partially miscible with the first solvent, mixing the two
dispersions and then either
removing the second or first solvent from the final mixture.
Alternatively, in some embodiments, the associating step may involve
dispersing both the
pre-formed polymer nanoparticles and active ingredient in a common solvent and
removing all or a
portion of the common solvent from the final mixture. The final form of the
nanoparticles of
Date Recue/Date Received 2020-06-05
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
polymer-associated active ingredient can be either a dispersion in a common
solvent or a dried solid.
The common solvent is typically one that is capable of swelling the polymer
nanoparticles as well as
dissolving the active ingredient at a concentration of at least about 10
mg/mL, e.g., at least about 20
mg/mL. The polymer nanoparticles are typically dispersed in the common solvent
at a concentration
of at least about 10 mg/mL, e.g., at least about 20 mg/mL. In some
embodiments, the common
solvent is an alcohol (either long or short chain), preferably methanol or
ethanol. In some
embodiments the common solvent is selected from alkenes, alkanes, alkynes,
phenols,
hydrocarbons, chlorinated hydrocarbons, ketones, and ethers. In some
embodiments, the common
solvent is a mixture of two or more different solvents that are miscible or
partially miscible with each
other. Some or all of the common solvent is removed from the dispersion of pre-
formed polymer
nanoparticles and active ingredients by either direct evaporation or
evaporation under reduced
pressure. The dispersion can be dried by a range of processes known by a
practitioner of the art
such as lyophilization (freeze-drying), spray-drying, tray-drying,
evaporation, jet drying, or other
methods to obtain the nanoparticles of polymers-associated with active
ingredients. In general, the
amount of solvent that is removed from the dispersion described above will
depend on the final type
of formulation that is desired. This is illustrated further in the Examples
and in the general
description of specific formulations.
In some instances the solids content (including both the polymer and active
ingredient
components as well as other solid form formulating agents) of the formulation
is between about 1
and about 98 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about land about 90 weight % of the total formulation.
In some
embodiments, the solids content of the formulation is between about 1 and
about 75 weight % of
the total formulation. In some embodiments, the solids content of the
formulation is between about
1 and about 50 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about land about 30 weight % of the total formulation.
In some
embodiments, the solids content of the formulation is between about 1 and
about 25 weight % of
the total formulation. In some embodiments, the solids content of the
formulation is between about
1 and about 10 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about 10 and about 25 weight % of the total
formulation. In some
embodiments, the solids content of the formulation is between about 10 and
about 30 weight % of
the total formulation. In some embodiments, the solids content of the
formulation is between about
10 and about 50 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about 10 and about 75 weight % of the total
formulation. In some
embodiments, the solids content of the formulation is between about 10 and
about 90 weight % of
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
31
the total formulation. In some embodiments, the solids content of the
formulation is between about
and about 98 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about 25 and about 50 weight % of the total
formulation. In some
embodiments, the solids content of the formulation is between about 25 and
about 75 weight % of
the total formulation. In some embodiments, the solids content of the
formulation is between about
25 and about 90 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about 30 and about 98 weight % of the total
formulation. In some
embodiments, the solids content of the formulation is between about 50 and
about 90 weight % of
the total formulation. In some embodiments, the solids content of the
formulation is between about
50 and about 98 weight % of the total formulation. In some embodiments, the
solids content of the
formulation is between about 75 and about 90 weight % of the total
formulation. In some
embodiments, the solids content of the formulation is between about 75 and
about 98 weight % of
the total formulation.
Formulating Agents
As used herein, the term "formulating agent" refers to any other material used
in the
formulation other than the nanoparticles of polymer-associated active
ingredient. Formulating
agents can include, but are not limited to, compounds that can act as a
dispersants or wetting
agents, inert fillers, solvents, surfactants, anti-freezing agents, anti-
settling agents or thickeners,
disintegrants, and preservatives.
In some embodiments, a formulation may include a dispersant or wetting agent
or both. In
some embodiments the same compound may act as both a dispersant and a wetting
agent. A
dispersant is a compound that helps the nanoparticles (or aggregates of
nanoparticles) disperse in
water. Without wishing to be bound by any theory, dispersants are thought to
achieve this result by
absorbing on to the surface of the nanoparticles and thereby limiting re-
aggregation. Wetting
agents increase the spreading or penetration power of a liquid when placed
onto the substrate (e.g.,
leaf). Without wishing to be bound by any theory, wetting agents are thought
to achieve this result
by reducing the interfacial tension between the liquid and the substrate
surface.
In a similar manner, some formulating agents may demonstrate multiple
functionality. The
categories and listings of specific agents below are not mutually exclusive.
For example, fumed
silica, described below in the thickener! anti-settling agent and anti-caking
agent sections, is
typically used for these functions. In some embodiments, however, fumed silica
demonstrates the
functionality of a wetting agent and/or dispersant. Specific formulating
agents listed below are
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
32
categorized based on their primary functionality, however, it is to be
understood that particular
formulating agents may exhibit multiple functions. Certain formulation
ingredients display multiple
functionalities and synergies with other formulating agents and may
demonstrate superior
properties in a particular formulation but not in another formulation.
In some embodiments, a dispersant or wetting agent is selected from
organosilicones (e.g.,
SYLGARD 309 from Dow Corning Corporation or SILWET L77 from Union Carbide
Corporation)
including polyalkylene oxide modified polydinnethylsiloxane (SILWET L7607 from
Union Carbide
Corporation), methylated seed oil, and ethylated seed oil (e.g., SCOIL from
Agsco or HASTEN from
Wilfarnn), alkylpolyoxyethylene ethers (e.g., ACTIVATOR 90), alkylarylalolates
(e.g., APSA 20),
alkylphenol ethoxylate and alcohol alkoxylate surfactants (e.g., products sold
by Huntsman), fatty
acid, fatty ester and fatty amine ethoxylates (e.g., products sold by
Huntsman), products sold by
Cognis such as sorbitan and ethoxylated sorbitan esters, ethoxylated vegetable
oils, alkyl, glycol and
glycerol esters and glycol ethers, tristyrylphenol ethoxylates, anionic
surfactants such as sulfonates,
such as sulfosuccinates, alkylaryl sulphonates, alkyl napthalene sulfonates
(e.g., products sold by
Adjuvants Unlimited), calcium alkyl benzene sulphonates, and phosphate esters
(e.g., products sold
by Huntsman Chemical or BASF), as salts of sodium, potassium, ammonium,
magnesium,
triethanolannine (TEA), etc.
Other specific examples of the above sulfates include ammonium lauryl sulfate,
magnesium
lauryl sulfate, sodium 2-ethyl-hexyl sulfate, sodium actyl sulfate, sodium
ley' sulfate, sodium
tridecyl sulfate, triethanolamine lauryl sulfate, ammonium linear alcohol,
ether sulfate ammonium
nonylphenol ether sulfate, and ammonium nnonoxyno1-4-sulfate. Other examples
of dispersants and
wetting agents include, sulfo succinamates, disodium N-octadecylsulfo-
succinamate; tetrasodium N-
(1,2-dicarboxyethyl)-N-octadecylsulfo-succinannate; diamyl ester of sodium
sulfosuccinic acid;
dihexyl ester of sodium sulfosuccinic acid; and dioctyl esters of sodium
sulfosuccinic acid; dihexyl
ester of sodium sulfosuccinic acid; and dioctyl esters of sodium sulfosuccinic
acid; castor oil and fatty
amine ethoxylates, including sodium, potassium, magnesium or ammonium salts
thereof.
Dispersants and wetting agents also include natural emulsifiers, such as
lecithin, fatty acids
(including sodium, potassium or ammonium salts thereof) and ethanolamides and
glycerides of fatty
acids, such as coconut diethanolannide and coconut mono- and diglycerides.
Dispersants and
wetting agents also include sodium polycarboxylate (commercially available as
Geropon TA/72);
sodium salt of naphthalene sulfonate condensate (commercially available as
Morwet (D425, D809,
D390, EFW); calcium naphthalene sulfonates (commercially available as DAXAD
19LCAD); sodium
lignosulfonates and modified sodium lignosulfonates; aliphatic alcohol
ethoxylates; ethoxylated
tridecyl alcohols (commercially available as Rhodasurf (BC420, BC610, BC720,
BC 840); Ethoxylated
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
33
tristeryl phenols (commercially available as Soprophor BSU); sodium methyl
oleyl taurate
(commercially available as Geropon T-77); tristyrylphenol ethoxylates and
esters; ethylene oxide-
propylene oxide block copolymers; non-ionic copolymers (e.g., commercially
available Atlox 4913),
non-ionic block copolymers (commercially available as Atlox 4912). Examples of
dispersants and
wetting agents include, but are not limited to, sodium dodecylbenzene
sulfonate; N-oleyl N-methyl
taurate; 1,4-dioctoxy-1,4-dioxo-butane-2-sulfonic acid; sodium lauryl
sulphate; sodium dioctyl
sulphosuccinate; aliphatic alcohol ethoxylates; nonylphenol ethoxylates.
Dispersants and wetting
agents also include sodium taurates; and sodium or ammonium salts of nnaleic
anhydride
copolymers, lignosulfonic acid formulations or condensed sulfonate sodium,
potassium, magnesium
or ammonium salts, polyvinylpyrrolidone (available commercially as
POLYPLASDONE XL-10 from
International Specialty Products or as KOLLI DON Cl M-10 from BASF
Corporation), polyvinyl
alcohols, modified or unmodified starches, methylcellulose, hydroxyethyl or
hydroxypropyl
methylcellulose, carboxymethyl methylcellulose, or combinations, such as a
mixture of either
lignosulfonic acid formulations or condensed sulfonate sodium, potassium,
magnesium or
ammonium salts with polyvinylpyrrolidone (PVP).
In some embodiments, the dispersants and wetting agents can combine to make up
between about 0.5 and about 30 weight % of the formulation. For example,
dispersants and wetting
agents can make up between about 0.5 and about 20 weight %, about 0.5 and
about 10 weight %,
between about 0.5 and about 5 weight %, between about 0.5 and about 3 weight
%, between about
1 and about 30 weight %, between about 1 and about 20 weight %, between about
1 and about 10
weight %, between about 1 and about 5 weight %, between about 2 and about 30
weight %,
between about 2 and about 20 weight %, between about 2 and about 10 weight %,
between about 2
and about 5 weight %, between about 3 and about 30 weight %, between about 3
and about 20
weight %, between about 3 and about 10 weight %, between about 3 and about 5
weight %,
between about 5 and about 30 weight %, between about 5 and about 20 weight %,
between about 5
and about 10 weight % of the formulation. In some embodiments, dispersants or
wetting agents can
make up between about 0.1 and 1 weight % of the formulation, between about 0.1
and 2 weight %
of the formulation between about 0.1 and 3 weight % of the formulation between
about 0.1 and 5
weight % of the formulation, between about 0.1 and 10 weight % of the
formulation.
In some embodiments, a formulation may include an inert filler. For example,
an inert filler
may be included to produce or promote cohesion in forming a wettable granule
formulation. An
inert filler may also be included to give the formulation a certain active
loading, density, or other
similar physical properties. Non limiting examples of inert fillers that may
be used in a formulation
include bentonite clay, carbohydrates, proteins, lipids synthetic polymers,
glycolipids, glycoproteins,
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
34
lipoproteins, lignin, lignin derivatives, and combinations thereof. In a
preferred embodiment the
inert filler is a lignin derivative and is optionally calcium lignosulfonate.
In some embodiments, the
inert filler is selected from the group consisting of: monosaccharides,
disaccharides,
oligosaccharides, polysaccharides and combinations thereof. Specific
carbohydrate inert fillers
illustratively include glucose, mannose, fructose, galactose, sucrose,
lactose, maltose, xylose,
arabinose, trehalose and mixtures thereof such as corn syrup; sugar alcohols
including: sorbitol,
xylitol , ribitol, mannitol, galactitol, fucitol, iditol, inositol, volemitol,
isomalt, maltitol, lactitol,
polyglycitol; cellu loses such as carboxynnethylcellulose, ethylcellulose,
hydroxyethylcellulose,
hydroxy-methylethylcellulose, hydroxyethylpropylcellulose,
methylhydroxyethylcellulose,
nnethylcellulose; starches such as annylose, seagel, starch acetates, starch
hydroxyethyl ethers, ionic
starches, long-chain alkyl starches, dextrins, amine starches, phosphates
starches, and dialdehyde
starches; plant starches such as corn starch and potato starch; other
carbohydrates such as pectin,
amylopectin, xylan, glycogen, agar, alginic acid, phycocolloids, chitin, gum
arabic, guar gum, gum
karaya, gum tragacanth and locust bean gum; vegetable oils such as corn,
soybean, peanut, canola,
olive and cotton seed; complex organic substances such as lignin and
nitrolignin; derivatives of lignin
such as lignosulfonate salts illustratively including calcium lignosulfonate
and sodium lignosulfonate
and complex carbohydrate-based formulations containing organic and inorganic
ingredients such as
molasses. Suitable protein inert fillers illustratively include soy extract,
zein, protamine, collagen,
and casein. Inert fillers operative herein also include synthetic organic
polymers capable of
promoting or producing cohesion of particle components and such inert fillers
illustratively include
ethylene oxide polymers, polyacrylamides, polyacrylates, polyvinyl
pyrrolidone, polyethylene glycol,
polyvinyl alcohol, polyvinylnnethyl ether, polyvinyl acrylates, polylactic
acid, and latex.
In some embodiments, a formulation contains between about 1 and about 90
weight % inert
filler, e.g., between about 1 and about 80 weight %, between about 1 and about
60 weight %,
between about 1 and about 40 weight %, between about 1 and about 25 weight %,
between about 1
and about 10 weight %, between about 10 and about 90 weight %, between about
10 and about 80
weight %, between about 10 and about 60 weight %, between about 10 and about
40 weight %,
between about 10 and about 25 weight %, between about 25 and about 90 weight
%, between
about 25 and about 80 weight %, between about 25 and about 60 weight %,
between about 25 and
about 40 weight %, between about 40 and about 90 weight %, between about 40
and about 80
weight %, or between about 60 and about 90 weight %.
In some embodiments, a formulation may include a solvent or a mixture of
solvents that can
be used to assist in controlling the solubility of the active ingredient
itself, the nanoparticles of
polymer-associated active ingredients, or other components of the formulation.
For example, the
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
solvent can be chosen from water, alcohols, alkenes, alkanes, alkynes,
phenols, hydrocarbons,
chlorinated hydrocarbons, ketones, ethers, and mixtures thereof. In some
embodiments, the
formulation contains a solvent or a mixture of solvents that makes up about
0.1 to about 90 weight
% of the formulation. In some embodiments, a formulation contains between
about 0.1 and about
90 weight % solvent, e.g., between about 1 and about 80 weight %, between
about 1 and about 60
weight %, between about 1 and about 40 weight %, between about 1 and about 25
weight %,
between about 1 and about 10 weight %, between about 10 and about 90 weight %,
between about
10 and about 80 weight %, between about 10 and about 60 weight %, between
about 10 and about
weight %, between about 10 and about 25 weight %, between about 25 and about
90 weight %,
between about 25 and about 80 weight %, between about 25 and about 60 weight
%, between
about 25 and about 40 weight %, between about 40 and about 90 weight %,
between about 40 and
about 80 weight %, between about 60 and about 90 weight %, between about 0.1
and about 10
weight %, between about 0.1 and about 5 weight %, between about 0.1 and about
3 weight %,
between about 0.1 and about 1 weight %, between about 0.5 and about 20 weight
%, 0 between
about.5 and about 10 weight %, between about 0.5 and about 5 weight %, between
about 0.5 and
about 3 weight %, between about 0.5 and about 1 weight %, between about 1 and
about 20 weight
%, between about 1 and about 10 weight %, between about 1 and about 5 weight
%, between about
1 and about 3 weight %, between about 5 and about 20 weight %, between about 5
and about 10
weight %, between about 10 or about 20 weight %.
In some embodiments, a formulation may include a surfactant. When included in
formulations, surfactants can function as wetting agents, dispersants,
emulsifying agents, solublizing
agents and bioenhancing agents. Without limitation, particular surfactants may
be anionic
surfactants, cationic surfactants, nonionic surfactants, annphoteric
surfactants, silicone surfactants
(e.g., Silwet L77), and fluorosurfactants. Exemplary anionic surfactants
include alkylbenzene
sulfonates, olefinic sulfonate salts, alkyl sulfonates and ethoxylates,
sulfosuccinates, phosphate
esters, taurates, alkylnaphthalene sulfonates and polymers lignosulfonates.
Exemplary nonionic
surfactants include alkylphenol ethoxylates, aliphatic alcohol ethoxylates,
aliphatic alkylannine
ethoxylates, amine alkoxylates, sorbitan esters and their ethoxylates, castor
oil ethoxylates, ethylene
oxide/propylene oxide copolymers and polymeric surfactants, non-ionic
copolymers (e.g.,
commercially available Atlox 4913), non-ionic block copolymers (commercially
available as Atlox
4912). In some embodiments, surfactants can make up between about 0.1 and
about 20 weight %
of the formulation, e.g., between about 0.1-15 weight %, between about 0.1 and
about 10 weight %,
between about 0.1 and about 8 weight %, between about 0.1 and about 6 weight
%, between about
0.1 and about 4 weight %, between about 1-15 weight %, between about 1 and
about 10 weight %,
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
36
between about 1 and about 8 weight %, between about 1 and about 6 weight %,
between about 1
and about 4 weight %, between about 3 and about 20 weight %, between about 3
and about 15
weight %, between about 3 and about 10 weight %, between about 3 and about 8
weight %,
between about 3 and about 6 weight %, between about 5 and about 15 weight %,
between about
and about 10 weight %, between about 5 and about 8 weight %, or between about
10 and about 15
weight %. In some embodiments, a surfactant (e.g., a non-ionic surfactant) may
be added to a
formulation by the end user, e.g., in a spray tank. Indeed, when a formulation
is added to the spray
tank it becomes diluted and, in some embodiments, it may be advantageous to
add additional
surfactant in order to maintain the nanoparticles in dispersed form.
Suitable non-ionic surfactants also include alkyl polyglucosides (APGs). Alkyl
polyglucosides
which can be used in the adjuvant composition herein include those
corresponding to the formula:
R40(R50)b(Z3) wherein R4 is a monovalent organic radical of from 6 to 30
carbon atoms; Rs is a
divalent alkylene radical of from 2 to 4 carbon atoms; Z3 is a saccharide
residue of 5 or 6 carbon
atoms; a is a number ranging from 1 to 6; and, b is a number ranging from 0 to
12. More specifically
R4 is a linear C6 to C12 group, b is 0, Z3 is a glucose residue, and a is 2.
Some non-limiting examples
of commercially available alkyl polyglucosides include, e.g., APGTM,
AgniqueTM, or AgrimulTM
surfactants from Cognis Corporation (now owned by BASF), and AGTM series
surfactants from Akzo
Nobel Surface Chemistry, LLC.
In some embodiments, a formulation may include an anti-settling agent or
thickener that
can help provide stability to a liquid formulation or modify the rheology of
the formulation.
Examples of anti-settling agents or thickeners include, but are not limited
to, guar gum; locust bean
gum; xanthan gum; carrageenan; alginates; methyl cellulose; sodium
carboxymethyl cellulose;
hydroxyethyl cellulose; modified starches; polysaccharides and other modified
polysaccharides;
polyvinyl alcohol; glycerol alkyd resins such as Latron B-1956 from Rohm &
Haas Co., plant oil based
materials (e.g., cocodithalymide) with emulsifiers; polymeric terpenes;
microcrystalline cellulose;
methacrylates; poly(vinylpyrrolidone), syrups, polyethylene oxide, hydrophobic
silica, hydrated silica
and fumed silica (e.g., Aerosil 380). In some embodiments, anti-settling
agents or thickeners can
make up between about 0.05 and about 10 weight % of the formulation, e.g.,
about 0.05 to about 5
weight %, about 0.05 to about 3 weight %, about 0.05 to about 1 weight %,
about 0.05 to about 0.5
weight %, about 0.05 to about 0.1 weight %, about 0.1 to about 5 weight %,
about 0.1 to about 3
weight %, about 0.1 to about 2 weight %, about 0.1 to about 1 weight %, about
0.1 to about 0.5
weight %, about 0.5 to about 5 weight %, about 0.5 to about 3 weight %, about
0.5 to about 1 weight
%, about 1 to about 10 weight %, about 1 to about 5 weight %, or about 1 to
about 3 weight %. In
some embodiments, it is explicitly contemplated that a formulation of the
present disclosure does
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
37
not include a compound whose primary function is to act as an anti-settling or
thickener. In some
embodiments, compounds included in a formulation may have some anti-settling
or thickening
functionality, in addition to other, primary functionality, so anti-settling
or thickening functionality is
not a necessary condition for exclusion, however, formulation agents used
primarily or exclusively as
anti-settling agents or thickeners may be expressly omitted from the
formulations.
In some embodiments, a formulation may include one or more preservatives that
prevent
microbial or fungal degradation of the product during storage. Examples of
preservatives include
but are not limited to, tocopherol, ascorbyl palnnitate, propyl gallate,
butylated hydroxyanisole
(BHA), butylated hydroxytoluene (BHT), propionic acid and its sodium salt;
sorbic acid and its sodium
or potassium salts; benzoic acid and its sodium salt; p-hydroxy benzoic acid
sodium salt; methyl p-
hydroxy benzoate; 1,2-benzisothiazalin-3-one, and combinations thereof. In
some embodiments,
preservatives can make up about 0.01 to about 0.2 weight % of the formulation,
e.g., between about
0.01 and about 0.1 weight %, between about 0.01 and about 0.05 weight %,
between about 0.01 and
about 0.02 weight %, between about 0.02 and about 0.2 weight %, between about
0.02 and about
0.1 weight %, between about 0.02 and about 0.05 weight %, between about 0.05
and about 0.2
weight %, between about 0.05 and about 0.1 weight %, or between about 0.1 and
about 0.2 weight
%.
In some embodiments, a formulation may include anti-freezing agents, anti-
foaming agents,
and/or anti-caking agents that help stabilize the formulation against freezing
during storage,
foaming during use, or caking during storage. Examples of anti-freezing agents
include, but are not
limited to, ethylene glycol, propylene glycol, and urea. In certain embodiment
a formulation may
include between about 0.5 and about 10 weight % anti-freezing agents, e.g.,
between about 0.5 and
about 5 weight %, between about 0.5 and about 3 weight %, between about 0.5
and about 2 weight
%, between about 0.5 and about 1_ weight %, between about 1 and about 10
weight %, between
about 1 and about 5 weight %, between about 1 and about 3 weight %, between
about 1 and about
2 weight %, between about 2 and about 10 weight %, between about 3 and about
10 weight %, or
between about 5 and about 10 weight %.
Examples of anti-foaming agents include, but are not limited to, silicone
based anti-foaming
agents (e.g., aqueous emulsions of dimethyl polysiloxane, FG-10 from Dow-
Corning, Trans 10A from
Trans-Chenno Inc.), and non-silicone based anti-foaming agents such as
octanol, nonanol, and silica.
In some embodiments a formulation may include between about 0.05 and about 5
weight % of anti-
foaming agents, e.g., between about 0.05 and about 0.5 weight %, between about
0.05 and about 1
weight %õ between about 0.05 and about 0.2 weight %, between about 0.1 and
about 0.2 weight %,
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
38
between about 0.1 and about 0.5 weight %, between about 0.1 and about 1 weight
%, or between
about 0.2 and about 1 weight %.
Examples of anti-caking agents include sodium or ammonium phosphates, sodium
carbonate
or bicarbonate, sodium acetate, sodium nnetasilicate, magnesium or zinc
sulfates, magnesium
hydroxide (all optionally as hydrates), sodium alkylsulfosuccinates, silicious
compounds, magnesium
compounds, C10 -C22 fatty acid polyvalent metal salt compounds, and the like.
Illustrative of anti-
caking ingredients are attapulgite clay, kieselguhr, silica aerogel, silica
xerogel, perlite, talc,
vermiculite, sodium aluminosilicate, aluminosilicate clays (e.g.,
Montnnorillonite, Attapulgite, etc.,)
zirconium oxychloride, starch, sodium or potassium phthalate, calcium
silicate, calcium phosphate,
calcium nitride, aluminum nitride, copper oxide, magnesium aluminum silicate,
magnesium
carbonate, magnesium silicate, magnesium nitride, magnesium phosphate,
magnesium oxide,
magnesium nitrate, magnesium sulfate, magnesium chloride, and the magnesium
and aluminum
salts of C10 -C22 fatty acids such as palmitic acid, stearic acid and oleic
acid. Anti-caking agents also
include refined kaolin clay, amorphous precipitated silica dioxide, such as HI
SIL 233 available from
PPG Industries, refined clay, such as HUBERSIL available from Huber Chemical
Company, or fumed
silica (e.g., Aerosil 380) In some embodiments, a formulation may include
between about 0.05 and
about 10 weight % anti-caking agents, e.g., between about 0.05 to 5 weight %,
between about 0.05
and about 3 weight %, between about 0.05 and about 2 weight %, between about
0.05 and about 1
weight %, between about 0.05 and about 0.5 weight %, between about 0.05 and
about 0.1 weight %,
between about 0.1 and about 5 weight %, between about 0.1 and about 3 weight
%, between about
0.1 and about 2 weight %, between about 0.1 and about 1 weight %, between
about 0.1 and about
0.5 weight %, between about 0.5 and about 5 weight %, between about 0.5 and
about 3 weight %,
between about 0.5 and about 2 weight %, between about 0.5 and about 1 weight
%, between about
1 to 3 weight %, between about 1 to 10 weight %, or between about 1 and about
5 weight %.
In some embodiments, a formulation may include a UV-blocking compound that can
help
protect the active ingredient from degradation due to UV irradiation. Examples
of UV-blocking
compounds include ingredients commonly found in sunscreens such as
benzophenones,
benzotriazoles, homosalates, alkyl cinnamates, salicylates such as octyl
salicylate,
dibenzoylnnethanes, anthranilates, methylbenzylidenes, octyl triazones, 2-
phenylbenzimidazole-5-
sulfonic acid, octocrylene, triazines, cinna mates, cyanoacrylates, dicyano
ethylenes, etocrilene,
dronnetrizole trisiloxane, bisethylhexyloxyphenol methoxyphenol triazine,
dronnetrizole, dioctyl
butamido triazone, terephthalylidene dicamphor sulfonic acid and para-
aminobenzoates as well as
ester derivatives thereof, UV-absorbing metal oxides such as titanium dioxide,
zinc oxide, and cerium
oxide, and nickel organic compounds such as nickel bis (octylphenol) sulfide,
etc. Additional
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
39
examples of each of these classes of UV-blockers may be found in Kirk-Othmer,
Encyclopedia of
Chemical Technology. In some embodiments, a formulation may include between
about 0.01 and
about 2 weight % UV-blockers, e.g., between about 0.01 and about 1 weight %,
between about 0.01
and about 0.5 weight %, between about 0.01 and about 0.2 weight %, between
about 0.01 and
about 0.1 weight %, between about 0.01 and about 0.05 weight %, between about
0.05 weight %
and about 1 weight %, between about 0.05 and about 0.5 weight %, between about
0.05 and about
0.2 weight %, between about 0.05 and about 0.1 weight %, between about 0.1 and
about 1 weight
%, between about 0.1 and about 0.5 weight %, between about 0.1 and about 0.2
weight %, between
about 0.2 and about 1 weight %, between about 0.2 and about 0.5 weight %, or
between about 0.5
and about 1 weight %. In some embodiments, it is explicitly contemplated that
a formulation of the
present disclosure does not include a compound whose primary function is to
act as a UV-blocker.
In some embodiments, compounds included in a formulation may have some UV-
blocking
functionality, in addition to other, primary functionality, so UV-blocking is
not a necessary condition
for exclusion, however, formulation agents used primarily or exclusively as UV-
blockers may be
expressly omitted from the formulations.
In some embodiments, a formulation may include a disintegrant that can help a
solid
formulation break apart when added to water. Examples of suitable
disintegrants include cross-
linked polyvinyl pyrrolidone, modified cellulose gum, pregelatinized starch,
cornstarch, modified
corn starch (e.g., STARCH 1500) and sodium carboxynnethyl starch (e.g.,
EXPLOTAB or PRIMOJEL),
nnicrocrystalline cellulose, sodium starch glycolate, sodium carboxynnethyl
cellulose, carnnellose,
carnnellose calcium, carnnellose sodium, croscarnnellose sodium, carnnellose
calcium,
carboxymethylstarch sodium, low-substituted hydroxypropyl cellulose,
hydroxypropyl
nnethylcellulose, hydroxypropyl cellulose, soy polysaccharides (e.g.,
EMCOSOY), alkylcelullose,
hydroxyalkylcellulose, alginates (e.g., SATIALGINE), dextrans and
poly(alkylene oxide) and an
effervescent couple (e.g., citric or ascorbic acid plus bicarbonate), lactose,
anhydrous dibasic calcium
phosphate, dibasic calcium phosphate, magnesium alunninonnetasilicate,
synthesized hydrotalcite,
silicic anhydride and synthesized aluminum silicate. In some embodiments
disintegrants can make
up between about 1 and about 20 weight % of the formulation, e.g., between
about 1 and about 15
weight %, between about 1 and about 10 weight %, between about 1 and about 8
weight %,
between about 1 and about 6 weight %, between about 1 and about 4 weight %,
between about 3
and about 20 weight %, between about 3 and about 15 weight %, between about 3
and about 10
weight %, between about 3 and about 8 weight %, between about 3 and about 6
weight %, between
about 5 and about 15 weight %, between about 5 and about 10 weight %, between
about 5 and
about 8 weight %, or between about 10 and about 15 weight %.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
Formulations
As described above, the nanoparticles of polymer-associated active ingredient
can be
formulated into different types of formulations for different applications.
For example, the types of
formulations can include wettable granules, wettable powders, and high solid
liquid suspensions.
Furthermore, as discussed above, formulation agents can include, but are not
limited to dispersants,
wetting agents, surfactants, anti-settling agents or thickeners,
preservatives, anti-freezing agents,
anti-foaming agents, anti-caking agents, inert fillers, and UV-blockers.
In some embodiments, a dispersion of polymer nanoparticles and active
ingredient in a
common solvent is dried (e.g., spray dried) to form a solid containing
nanoparticles (optionally in
aggregate form) of polymer-associated active ingredients. The spray dried
solid can then be used as
is or incorporated into a formulation containing other formulating agents to
make a wettable
granule (WG), wettable powder (WP), or a high solids liquid suspension (HSLS).
In some embodiments, active ingredient is milled in the presence of pre-formed
polymer
nanoparticles to form a solid containing nanoparticles (optionally in
aggregate form) of polymer-
associated active ingredients. The solid can then be used as is or
incorporated into a formulation
containing other formulating agents to make a wettable granule (WG), wettable
powder (WP), or a
high solids liquid suspension (HSLS). In some embodiments, the milling step
may be performed in the
presence of one or more formulating agents. In some embodiments, the milling
step may be
performed in the presence of an aqueous phase.
Wettable Powder (WP)
In some embodiments, the dried solid can be made into a formulation that is a
wettable
powder (WP). In some embodiments, a WP formulation comprising nanoparticles of
polymer-
associated active ingredients (optionally in aggregate form) can be made from
a dried (e.g., spray
dried, freeze dried, etc.) dispersion of polymer nanoparticles and active
ingredient. In some
embodiments, a WP formulation comprising nanoparticles of polymer-associated
active ingredients
(optionally in aggregate form) can be made from a milled solid comprising
polymer nanoparticles of
active ingredient. In some embodiments, a WP is made by mixing the dried solid
with a dispersant
and/or a wetting agent. In some embodiments, a WP is made by mixing the dried
solid or milled
solid with a dispersant and/or a wetting agent. In some embodiments, a WP is
made by mixing the
dried or milled solid with a dispersant and a wetting agent. In some
embodiments, the formulation
of the final WP can be (by weight): up to about 98% nanoparticles of polymer-
associated active
ingredients (including both the active ingredient and the polymer, optionally
in aggregate form). In
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
41
some embodiments, the WP formulation includes (by weight): 0-5% dispersant, 0-
5% wetting agent,
5-98% nanoparticles of polymer-associated active ingredients (optionally in
aggregate form), and
inert filler to 100%. In some embodiments, the formulation of the final WP can
be (by weight): 0.5-
5% dispersant, 0.5%-5% wetting agent, 5-98% nanoparticles of polymer-
associated active ingredients
(optionally in aggregate form), and inert filler to 100%. As described above
in the Formulating
Agents and Nanoparticles of polymer-associated active ingredient sections, a
wide variety of
formulating agent(s) and various concentrations of nanoparticles (including
aggregates), wetting
agents, dispersants, fillers and other formulating agents can be used to
prepare exemplary
formulations, e.g. wettable granules.
In some embodiments, the formulation of the final WP can be (by weight): 0.5-
5%
dispersant, 0.5%-5% wetting agent, 0.1¨ 10% thickener (e.g., fumed silica
which, as noted above
may serve multiple functions, and/or xanthan gum), 5-98% nanoparticles of
polymer-associated
active ingredients (optionally in aggregate form). As described above in the
Formulating Agents
section, a wide variety of formulating agent(s) and various concentrations of
wetting agents,
dispersants, fillers and other formulating agents can be used to prepare
exemplary formulations, e.g.
wettable powders.
In some exemplary embodiments, described in more detail below, a WP
formulation
comprising nanoparticles of polymer-associated active ingredients (optionally
in aggregate form)
may be made from a dispersion of polymer nanoparticles and active ingredient
in a common solvent,
preferably methanol. In some embodiments, a WP formulation can be made by
adding the
dispersion in common solvent into an aqueous solution containing a wetting
agent (e.g., a surfactant
such as sodium dodecylbenzene sulfonate) and/or a dispersant (e.g., a
lignosulfonate such as Reax
888, etc.) and optionally an inert filler (e.g., lactose), and then drying
(e.g., freeze drying, spray
drying, etc.) the resulting mixture to from a solid powder. In some
embodiments, poly(vinyl alcohol)
is added to the solution prior to drying. In some embodiments a WP can be made
using a wetting
agent (e.g., a surfactant such as sodium dodecylbenzene sulfonate or dioctyl
sulfosuccinate sodium
salt) and a dispersant (e.g., a lignosulfonate such as Reax 8813, etc.).
In some exemplary embodiments, the polymer nanoparticles are made from a co-
polymer of
nnethacrylic acid and ethyl acrylate at about a 90:10 mass ratio. In some
embodiments, the polymer
nanoparticles are dispersed in a common solvent, preferably at a concentration
of about 50 nng/mL.
In some embodiments, the concentration of active ingredient is in the range
between about 20
ring/nnL to about 100 mg/nnl_. In some embodiments, the common solvent
contains a wetting agent
and/or dispersant as well. In some embodiments, the polymer nanoparticles are
made from a co-
polymer of nnethacrylic acid and (ethylene glycol)nnethyl ether nnethacrylate
at about at a mass ratio
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
42
of 7:3. In some embodiments, the polymer nanoparticles are made from a polymer
of acrylic acid.
In some embodiments, the polymer nanoparticles are made from a co-polymer of
acrylic acid and
styrene at about a 90:10 mass ratio. As described above in the Nanoparticles
of polymer-associated
active ingredient section, many ratios of co-polymer constituents can be used.
In some embodiments, the dispersion of polymer nanoparticles and active
ingredient is then
slowly added into a vessel containing a second solvent, preferably water. In
some embodiments, the
second solvent is at least 20 times larger in volume than the common solvent
containing the
polymer nanoparticles and active ingredient. In some embodiments, the second
solvent contains a
dispersant, preferably a lignosulfonate such as Reax 88B and/or a wetting
agent, preferably a
surfactant such as sodium dodecylbenzene sulfonate. In some embodiments a WP
can be made
using a wetting agent (e.g., a surfactant such as sodium dodecylbenzene
sulfonate or dioctyl
sulfosuccinate sodium salt) and a dispersant (e.g., a lignosulfonate such as
Reax 888, etc.).
In some embodiments, after the dispersion of polymer nanoparticles and active
ingredient in
a common solvent is mixed with a second solvent containing dispersant and/or
wetting agent, the
final mixture is dried (e.g., freeze dried) to obtain a solid powdered
formulation containing
nanoparticles of polymer-associated active ingredients (optionally in
aggregate form). Optionally,
the pH of the final mixture can be adjusted (e.g., by addition of acid or base
solutions) as needed.
Further, additional formulation agents (e.g., PVA solution) can also be added
to the final mixture
prior to drying.
High Solids Liquid Suspension (HSLS)
One type of formulation that can be utilized according to the disclosure is a
high solids liquid
suspension. As described, such a formulation is generally characterized in
that it is a liquid
formulation that contains at least nanoparticles of polymer nanoparticles
associated with active
ingredient (includes potentially aggregates of the same). HSLS formulations
most closely resemble
suspension concentrate (SC) formulations and can be considered a subcategory
SCs incorporating
polymer nanoparticles which are associated or encapsulate the active
ingredient and have a smaller
average particle size.
In some embodiments, the formulation of the HSLS can be (by weight): between
about 1 and
about 75% nanoparticles of polymer-associated active ingredients (including
both polymer and
active ingredient, optionally in aggregate form), 0.5 and about 5% wetting
agent and/or dispersant,
between about 1 and about 10% anti-freezing agent, between about 0.1 and about
10% anti-
foaming agent, between about 0.01 and about 0.1 % preservative, between about
0.1 and 4%
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
43
surfactant, and water up to 100% As described above in the Formulating Agents
and Nanoparticles
of polymer-associated active ingredient sections, a wide variety of
formulating agent(s) and various
concentrations of nanoparticles (including aggregates), wetting agents,
dispersants, fillers and other
formulating agents can be used to prepare exemplary formulations, e.g., a
HSLS.
In some exemplary embodiments, described in more detail below, the polymer
nanoparticles are made from a co-polymer of methacrylic acid and styrene at
about a 75:25 mass
ratio. In some exemplary embodiments, the polymer nanoparticles are dispersed
in the common
solvent, preferably at a concentration of up to about 20 mg/alL. In some
exemplary embodiments,
the active ingredient is difenoconazole and is mixed into the nanoparticle
dispersion at a
concentration of up to about 20 nng/rinL. As described above in the
Nanoparticles of polymer-
associated active ingredient section, many ratios of co-polymer constituents
can be used.
In some embodiments, the dispersion of polymer nanoparticles and active
ingredient in a
common solvent is slowly added into a vessel containing a second solvent,
preferably water. In
some embodiments, the second solvent is at least 20 times larger in volume
than the common
solvent containing the polymer nanoparticles and active ingredient. In some
embodiments, the
second solvent contains a dispersant, preferably a lignosulfonate such as Reax
88B and/or a wetting
agent, preferably a surfactant such as sodium dodecylbenzene sulfonate. In
some embodiments a
HSLS can be made using a wetting agent (e.g., a surfactant such as sodium
dodecylbenzene
sulfonate) and a dispersant (e.g., a lignosulfonate such as Reax 88B, etc.).
In some embodiments, the HSLS formulations of current disclosure have an
active ingredient
content of about 5 to about 40% by weight, e.g., about 5 ¨ about 40%, about 5
¨ about 35 %, about
¨ about 30%, about 5 ¨ about 25%, about 5 ¨ about 20%, about 5 ¨ about 15%,
about 5 ¨ about
10%, about 10¨ about 40%, about 10¨ about 35 %, about 10¨ about 30%, about 10¨
about 25%,
about 10¨ about 20%, about 10¨ about 15%, about 15 ¨ about 40%, about 15¨
about 35 %, about
15¨ about 30%, about 15¨ about 25%, about 15¨ about 20%, about 20¨ about 40%,
about 20 ¨
about 35%, about 20¨ about 30%, about 20¨ about 25 %, about 25¨ about 40%,
about 25 ¨ about
35%, about 25¨ about 30%, about 30¨ about 40% or about 35¨ about 40%. As
described above
in the Nanoparticles of polymer-associated active ingredient section, many
ratios of triazole to
polymer can be used.
In some embodiments the HSLS formulations of current disclosure have an active
ingredient
content of about 5 %, about 10%, about 15%, about 20%, about 25 %, about 30%,
about 35 % or
about 40% by weight.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
44
Methods of Making HSLS ¨Generally
In some embodiments, a HSLS comprising nanoparticles of polymer-associated
active
ingredient (optionally in aggregate form) can be made from a dispersion of
polymer nanoparticles
and active ingredient in a common solvent or from a dried form of the
dispersion (e.g., spray dried).
In some embodiments, a HSLS formulation comprising nanoparticles of polymer-
associated active
ingredients (optionally in aggregate form) can be made from a milled solid
comprising polymer
nanoparticles of active ingredient.
Methods of Making HSLS¨ Milling Methods
In some embodiments, a HSLS formulation comprising nanoparticles of polymer-
associated
active ingredients (optionally in aggregate form) can be prepared via milling.
Several exemplary
methods and the resulting HSLS formulations are described below and in the
Examples. In some
embodiments, a solid formulation of nanoparticles of polymer-associated active
ingredient
(optionally in aggregate form), prepared as described in this disclosure
(e.g., via milling, spray drying
etc.) may be further milled in the presence of one or more formulating agents
and water. In some
embodiments a HSLS can be made by milling a solid formulation nanoparticles of
polymer-associated
active ingredients in the presence of water and one more of an anti-freezing
agent, (optionally more
than one of) a wetter and/or dispersant, an antifoaming agent, a preservative,
and a thickening
agent. Further, in some embodiments, the active ingredient and polymer
nanoparticles are milled
together to produce nanoparticles of polymer-associated active ingredients,
which may then be
further milled according to the processes described below.
In some embodiments, the milling process is performed in separate phases
(i.e., periods of
time), with the optional addition of one or more formulating agent between
each milling phase. One
of ordinary skill in the art can adjust the length of each phase as is
appropriate for a particular
instance. In some embodiments, the contents of the milling vessel are cooled
between one or more
of milling phases (e.g., via placement of the milling jar in an ice bath). One
of ordinary skill in the art
can adjust the length of cooling period as is appropriate for a particular
instance.
In some embodiments, a HSLS can be made by first milling a solid formulation
of
nanoparticles of polymer-associated active ingredients in the presence of
(optionally more than one
of) a wetter and/or dispersant in one milling vessel for a certain amount of
time (e.g., about 30
minutes ¨ about 1 day), then this mixture is transferred to another milling
vessel containing water
and optionally one or more of an anti-freezing agent, additional wetter and/or
dispersant, an anti-
freezing agent, an antifoanning agent, a preservative, a thickening agent, and
milling the components
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
together. As described above in the Formulating Agents section, a wide variety
of additional
formulating agent(s) and various concentrations of wetting agents,
dispersants, fillers and other
formulating agents can be used in preparation of exemplary formulations.
In some embodiments, a HSLS formulation comprising nanoparticles of polymer-
associated
active ingredients (optionally in aggregate form) can be prepared via milling
pre-formed polymer
nanoparticles and active ingredient in the presence of one or more formulating
agents and water. In
some embodiments, a HSLS can be made by milling preformed polymer
nanoparticles and active
ingredient in the presence of water and optionally one more of an anti-
freezing agent, additional
wetter and/or dispersant, an anti-freezing agent, an antifoanning agent, a
preservative, and a
thickening agent. Again, as described above in the Formulating Agents section,
a wide variety of
additional formulating agent(s) and various concentrations of wetting agents,
dispersants, fillers and
other formulating agents can be used in preparation of exemplary formulations.
In some
embodiments, all of the ingredients can be added together and milled together.
And as in the embodiment described above in which nanoparticles of polymer-
associated
active ingredients are milled in a two milling vessel procedure, such a
procedure can be used in
preparing a HSLS from pre-formed polymer nanoparticles. In some embodiments
such an HSLS can
be made by first milling a solid formulation nanoparticles of polymer-
associated active ingredients in
the presence of (optionally more than one of) a wetter and/or dispersant in
one milling vessel for a
certain amount of time (e.g., about 30 minutes ¨ about 1 day), transferring
the milled components
to another milling vessel containing water and optionally one or more of an
anti-freezing agent,
additional wetter and/or dispersant, an anti-freezing agent, an antifoaming
agent, a preservative
and a thickening agent.
Milling methods to produce HSLS formulations as described above may include
any of those
referred to in any other portion of the specification including the Examples
below. Any type of mill
noted in any portion of the specification may also be used to prepare HSLS
formulations via milling.
Methods of Making HSLS¨Mixing & Drying Methods
In some embodiments, a HSLS formulation is prepared without milling, but
instead by mixing
the components of the formulation. These methods may also include drying the
formulations to
increase the solids content of the formulation so that it is suitable as a
HSLS. All of these methods
are described in more detail below and exemplary methods are shown in the
Examples.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
46
In some embodiments, a HSLS formulation comprising nanoparticles of polymer-
associated
active ingredients (optionally in aggregate form) can be made from the
dispersion of polymer
nanoparticles and active ingredient in a common solvent, (e.g., methanol). In
some embodiments,
the dispersion is added to an aqueous solution containing a wetting agent and
a dispersant, an anti-
freezing agent (and optionally an anti-settling agent or thickener and a
preservative). The mixture is
then concentrated by removing solvent, e.g., by drying, until the desired high
solids formulation is
attained.
In some exemplary embodiments, after the dispersion of polymer nanoparticles
and active
ingredient in a common solvent is mixed with a second solvent containing a
wetting agent and/or
dispersant and an anti-freezing agent (optionally with an anti-settling agent
or thickener and a
preservative), the final mixture is concentrated by removing most of the
common solvent and
second solvent until a final formulation with a target solids content (e.g.,
at least 60% solids) is
obtained. In some embodiments, the method used to concentrate the solution is
vacuum
evaporation. In some embodiments, a second solvent containing a wetting agent
and/or dispersant
and an anti-freezing agent (optionally with an anti-settling agent or
thickener and a preservative) are
added after the mixture has already been concentrated. As described above in
the Nanoparticles of
polymer-associated active ingredient section, many ranges of solids content
can be achieved.
In some embodiments, the dispersion of polymer nanoparticles and active
ingredient in a
common solvent is added to a second solvent to form a solution of
nanoparticles of polymer-
associated active ingredients (optionally in aggregate form). The second
solvent is typically miscible
with the common solvent and is usually water, but in some embodiments, the
second solvent can
also be a mixture of water with a third solvent, usually an alcohol,
preferably methanol or ethanol.
In some embodiments, the second solvent or mixture of solvents is only
partially miscible with the
common solvent. In some embodiments, the second solvent or mixture of solvents
is not miscible
with the common solvent. In some embodiments, the HSLS formulation is stable
after 1-2 months of
continuous temperature cycling between -5 C and 45 C showing no visible signs
of phase separation,
remains flowable, and can easily be dispersed in water at the use rate.
In some embodiments, a HSLS is made by reconstituting the dried dispersion
(e.g., freeze
dried) of nanoparticles of polymer-associated active ingredients in water to
obtain a formulation
with a target solids content (e.g., at least 60% solids) is obtained and then
adding an anti-freezing
agent (and optionally a thickening agent and a preservative) to the final
mixture. In some
embodiments, a HSLS is made by reconstituting the milled (e.g., ball-milled)
solid of nanoparticles of
polymer-associated active ingredients in water to obtain a formulation with a
target solids content
(e.g., at least 60% solids) and then adding an anti-freezing agent (and
optionally at least one
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
47
thickening agent (e.g., fumed silica and/or xanthan gum), an antifoaming agent
and a preservative)
to the final mixture. In some embodiments, the HSLS is made by homogenizing
all the components
together. In some embodiments the HSLS is made by milling all the components
together.
In some embodiments, a HSLS is made by mixing the dried dispersion (e.g.,
spray dried) with
a wetting agent, preferably a surfactant such as sodium dodecylbenzene
sulfonate, a solvent,
preferably but not limited to water, and/or a dispersant, preferably, but not
limited to a
lignosulfonate such as Reax 88B, and an anti-freezing agent, preferably but
not limited to ethylene
glycol, in a high sheer mixer until a stable HSLS is obtained. In some
embodiments a wetting agent,
preferably a surfactant such as sodium dodecylbenzene sulfonate, a solvent,
preferably but not
limited to water, and a dispersant, preferably, but not limited to a
lignosulfonate such as Reax 88B
are included. In some embodiments, a preservative, preferably propionic acid
and an anti-settling
agent or thickener, preferably but not limited to fumed silica and/or a water
dispersible agent like
xanthan gum are also included.
Efficacy and Application
General Applications and Efficacy
As noted previously and in the Examples, in some embodiments, the disclosure
provides
formulations of triazole compounds that have either improved curative,
translocation and/or
systemic fungicidal properties. In some embodiments, the triazole formulations
of the present
disclosure demonstrate improved preventative activity compared to commercial
formulations of the
same active ingredient, which suggests that they may be applied at lower
effective rates in
preventative applications. In some embodiments, the triazole formulations of
the present disclosure
demonstrate enhanced curative properties compared to commercial formulations
of the same active
ingredient, which suggests that they may be applied at lower effective rates
in curative applications.
Without wishing to be limited by any theory, it is thought that the enhanced
curative properties are
due to increased foliar penetration or translocation of triazoles formulated
according to the present
disclosure compared to triazoles of commercially available formulations. In
some embodiments, the
triazole formulations of the current disclosure can be applied at lower
effective rates than
commercial formulations for the control of fungal plant disease. In some
embodiments, the triazole
is difenoconazole.
In general, different triazoles are typically applied at different effective
rates between 10-
400 gram of active ingredient (e.g. triazole) per hectare depending on the
efficacy of the triazole
(e.g., absolute potency of the active and retention at the site of activity),
as well as conditions
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
48
related to the crop being treated, leaf type, environmental conditions, the
species infesting the crop,
infestation levels, and other factors. As discussed above, improvements in the
formulation
according to the current disclosure, such as increased UV stability, physical
retention at the site of
action, residual activity, systemic absorption, or enhanced curative activity
can reduce the user
rates. Some embodiments demonstrate improvements over typical commercial
formulation, which
suggests that lower rates of effective application could be used. In some
embodiments, rates may
range from between about 0.1 and about 400 g/hectare, preferably between about
0.1 and about
200 g/hectare, more preferably between about 0.1 and about 100 g/hectare, more
preferably
between about 0.1 and about 10g/hectare or more preferably between about 0.1
and about
1g/hectare. In some embodiments, rates may range from between about 1g and
about 400
g/hectare, preferably between about 1 and about 200 g/hectare, more preferably
between about 1
and about 100 g/hectare, or more preferably between about 1 and about 10
g/hectare. In some
embodiments, rates may be any of the rates or ranges of rates noted in any
other portion of the
specification.
General Application & Comparison to Current Commercial Formulations
In some embodiments, the disclosure provides methods of using formulations of
nanoparticles of polymer-associated triazoles. In some embodiments, the
formulations are used to
inoculate a target area of a plant. In some embodiments, the formulations are
used to inoculate a
part or several parts of the plant, e.g., the leaves, stem, roots, flowers,
bark, buds, shoots, and/or
sprouts.
In some embodiments, a formulation comprising nanoparticles of polymer-
associated active
ingredients and other formulating agents is added to water (e.g., in a spray
tank) to make a
dispersion that is about 10 to about 2,000 ppm in active ingredient. In some
embodiments, the
dispersion is about 10 to about 1,000 ppm, about 10 to about 500 ppm, about 10
to about 300 ppm,
about 10 to about 200 ppm, about 10 to about 100 ppm, about 10 to about 50
ppm, about 10 to
about 20 ppm, about 20 to about 2,000 ppm, about 20 to about 1,000 ppm, about
20 to about SOO
ppm, about 20 to about 300 ppm, about 20 to about 200 ppm, about 20 to about
100 ppm, about 20
to about SO ppm, about SO to about 2,000 ppm, about SO to about 1,000 ppm,
about SO to about
500 ppm, about 50 to about 300 ppm, about 50 to about 200 ppm, about 50 to
about 100 ppm,
about 100 to about 2,000 ppm, about 100 to about 1,000 ppm, about 100 to about
500 ppm, about
100 to about 300 ppm, about 100 to about 200 ppm, about 200 to about 2,000
ppm, about 200 to
about 1,000 ppm, about 200 to about 500 ppm, about 200 to about 300 ppm, about
300 to about
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
49
2,000 ppm, about 300 to about 1,000 ppm, about 300 to about 500 ppm, about 500
to about 2,000
ppm, about SOO to about 1,000 ppm, about 1000 to about 2,000 ppm.
As used in the specification, inoculation of a plant with a formulation of the
current
disclosure may, in some embodiments, refer to inoculation of a plant with a
dispersion (e.g., in water
or an aqueous medium optionally further comprising other additive such as
adjuvants, surfactants
etc.) prepared from a formulation of the present disclosure as described
above. It is to be
understood that the term formulation may also encompass dispersions for
applications as described
(e.g., inoculation of a plant). It should also be understood that methods that
describe the use of
triazole formulations of the present disclosure e.g., "use of formulations of
the present disclosure to
inoculate a plant," "use of the formulations of the present disclosure to
control fungal diseases" and
the like, encompass the preparation of a dispersion of the active ingredient
in water or an aqueous
medium (optionally further comprising other additives such as adjuvants,
surfactants etc.) for the
purpose of inoculating a plant.
In some embodiments, a dispersion is produced and used to inoculate a plant
with active
ingredient at less than about 75% of a use rate listed on a label of a
currently available commercial
product of the same active ingredient. In some embodiments, a dispersion is
produced to inoculate a
plant with active ingredient at less than about 60% of a use rate listed on
the label of a currently
available commercial product of the same active ingredient. In some
embodiments, a dispersion is
produced to inoculate a plant with active ingredient at less than about 50 %
of a use rate listed on
the label of a currently available commercial product of the same active
ingredient. In some
embodiments, a dispersion is produced to inoculate a plant with active
ingredient at less than 40%
of a use rate listed on the label of a currently available commercial product
of the same active
ingredient. In some embodiments, a dispersion is produced to inoculate a plant
with active
ingredient at less than 30% of a use rate listed on the label of a currently
available commercial
product of the same active ingredient. In some embodiments, a dispersion is
produced to inoculate a
plant with active ingredient at less than 25% of a use rate listed on the
label of a currently available
commercial product of the same active ingredient. In some embodiments, a
dispersion is produced
to inoculate a plant with active ingredient at less than 20% of a use rate
listed on the label of a
currently available commercial product of the same active ingredient. In some
embodiments, a
dispersion is produced to inoculate a plant with active ingredient at less
than 10% of a use rate
listed on the labels of a currently available commercial product of the same
active ingredient. In
some embodiments, a dispersion is produced to inoculate a plant with active
ingredient at less than
5% of the use rate listed on a label of a currently available commercial
product of the same active
ingredient. In some embodiments, the triazole formulations of the present
disclosure are used to
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
inoculate a plant at an active ingredient use rate that is about 75 %, about
60%, about 50 %, about
40%, about 30%, about 25%, about 20 % or about 10% of a use rate listed on the
labels of currently
available fungicide products. Fungicide labels can be referenced from
commercial suppliers and are
readily accessible and available.
In preferred embodiments, the formulations of the current disclosure may be
used to
control fungal disease at an active ingredient use rate that is lower than the
minimum rate of a
range of rates listed on the label of a commercially available product. In
some embodiments,
formulations of the current disclosure may be used to control fungal disease
at an active ingredient
use rate that is less than about 75%, less than about 60%, less than about 50
%, less than about 40
%, less than about 30%, less than about 25%, less than about 20% or less than
about 10 % of the
minimum use rate of a range of rates listed on the label of a commercially
available product.
Low Concentration Application
In some cases, a triazole formulation is applied to the plant at a
concentration below the
triazole's solubility limit in water. Although the active ingredient is
soluble in water at these low
concentrations, the triazole's activity is still affected by the way it is
formulated. This is surprising as
it demonstrates that the triazole is still associated with the polymer
particle even when applied
below its solubility limit. At concentrations below the solubility limits it
is expected that the triazoles
would behave the same, or at least in a very similar fashion, regardless of
the formulations,
especially with respect to biological functions described above. This is
because the triazoles are still
hydrophobic and thus, thought to still have low soil mobility, lack systemic
effects and display the
traits of traditional triazole and traditional triazole formulations.
In some embodiments, however, a formulation with nanoparticles or aggregates
of
nanoparticles of polymer associated triazole compound is shown to be more
active (e.g., have
systemic or curative effects) than commercially available suspension
concentrates of a triazole when
applied at a use rate below the solubility limit. Comparative example is
described below in the
Examples section. In some embodiments, the triazole is difenoconazole. In some
embodiments, the
polymer nanoparticles associated with the triazole compound is made from a
copolymer of
nnethacrylic acid and styrene at a mole ratio of 75:25 (MAA:S) though other
ratios and monomers,
as described above, are applicable. In some embodiments, the formulation
includes a wetter,
dispersant and filler.
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
51
Hard Water! Fertilizer Applications
As described below, most traditional formulations produce solid particles
(floc) or a
precipitate when mixed in with high salt, hard water or fertilizer solutions.
Surprisingly, a dispersed
solid formulation of a triazole (e.g., difenoconazole) of the current
disclosure was stable (e.g.,
components, difenoconazole and the salt, remained disperse, i.e., no visible
sedimentation or floc)
when mixed with a concentrated/high salt solution (e.g., hard water, buffer,
concentrated fertilizer
formulation) for at least 3 hours. This was true even for waters with ionic
strength as high as 8000
ppm Me (a.k.a. CIPAC "G" hard water). It is important to note that for such a
mixture to be useful
for the end user, the mixture should remain stable (i.e., no formation of
sediments and/or flocs)
within at least about 30 ¨ 40 minutes ¨ which is typically the time it takes
for the mixture to be
applied to the plant. It is surprising that the formulations of the present
disclosure are stable in such
high-salt conditions. Because the polymers that are used in the nanoparticles
of the present
disclosure are negatively charged, a practitioner of the art would expect the
formulations of the
present disclosure to flocculate when mixed with such a high amount of
divalent salt. Without being
limited by theory, it is believed that the increased stability of the
formulations of the present
disclosure arises from the use of nanoparticulate polymers as the delivery
system and that if
standard non-nanoparticle polymers were used then flocculation would occur
Traditional solid or liquid formulations are not stable under conditions of
high ionic (i.e., a
high salt solution) strength. Sources of increased ionic strength can include,
for example, mineral
ions that are present in the water that a formulation is dispersed in. For
example, in many cases the
water that is available to a farmer is taken from a high-salt ("hard water")
source such as a well or
aquifer. Water that a grower uses can be variably hard and is normally
measured as Ca2+
equivalents. Ranges of water salinity can be from ¨0 ppm Ca2F equivalent
(deionized water) to 8000
ppm Ca2+ or more.
Other sources of increased ionic strength can include, for example, other
chemicals or
materials that dispersed in the spray tank water before or after the addition
of the fungicide
formulation. Examples of this include mineral additives such as
nnicronutrients (which can include
e.g., B, Cu, Mn, Fe, CI, Mo, Zn, 5) or traditional N-P-K fertilizers where the
nitrogen, phosphorus, or
potassium source is in an ionic form as well as other agro-chemicals (e.g.,
pesticides, herbicides,
etc.). In some embodiments, the fertilizer can be, for example, 10-34-0 (N-P-
K), optionally including
one or more of sulfur, boron and another micronutrient. In some cases, the
nitrogen source is in the
form of urea or an agriculturally acceptable urea salt. The fertilizer can
include e.g., ammonium
phosphate or ammonium thiosulphate.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
52
In some embodiments described below in the Examples, the formulations of the
current
disclosure were mixed with a concentrated/high salt solution. Though the
specifics of the hard test
are described in Examples below, generally, the exemplary procedure is as
follows: Formulations
described herein were mixed with different hard water standards, each with a
different degree of
hardness (e.g., CIPAC H standard water (in the example below: 634 ppm
hardness, pH 6.0 ¨ 7.0, Ca2+:
= 2.5:1), CIPAC J standard water (6.34 ppm hardness, pH 6.0 ¨ 7.0, Ca2F: Mg2F
= 2.5:1) and CIPAC
G standard water (8000 ppm hardness, pH 6.0 ¨ 7.0, Mg2+)) at an active
ingredient concentration of
200 ppm. In some embodiments, the formulations dispersed well and were stable
for at least an
hour, with no signs of the formation of flocs or sediments.
In some cases, the formulations of the present disclosure can be applied
simultaneously with
a high-salt solution or suspension such as a nnicronutrient solution, a
fertilizer, pesticide, herbicide
solution, or suspension (e.g., in furrow application). The ability to mix and
apply triazoles with other
agricultural ingredients such as liquid fertilizers is very useful to growers,
as it reduces the number of
required trips across crop fields and the expenditure of resources for
application. In some cases, the
formulations of the present disclosure may be mixed with liquid fertilizers of
high ionic strength. In
some cases the fertilizer is a 10-34-0 fertilizer, optionally including one or
more of sulfur, boron and
another nnicronutrient. In some cases, the nitrogen source is in the form of
urea or an agriculturally
acceptable urea salt. In some embodiments, the liquid fertilizer comprises a
glyphosate or an
agriculturally acceptable salt of glyphosate (e.g., ammonium, isopropylannine,
dinnethylamine or
potassium salt). In some embodiments, the liquid fertilizer may be in the form
of a solution or a
suspension. In some embodiments, formulations of the present disclosure are
stable when mixed
with liquid fertilizers of increased or high ionic strength (e.g., at any of
the ionic strengths described
below). In some embodiments, when mixed with liquid fertilizers formulations
of the current
disclosure show no signs of sedimentation or flocculation. In some
embodiments, the triazole is
difenoconazole.
Other potential additives that might be added into a spray tank that are
charged and can
decrease the stability of an agrochemical formulation include charged
surfactants or polymers, inert
ingredients such as urea, or other similar ingredients.
In some embodiments, the present disclosure provides compositions of a
formulation of
nanoparticles of polymer-associated active ingredients that are redispersible
in solutions with high
ionic strength. In some embodiments, the present disclosure also provides
compositions of a
formulation of nanoparticles of polymer-associated active ingredients that can
be redispersed in
water and then have a high salt solution or solid salt added and maintain
their stability. In some
embodiments, the formulations of the present disclosure are stable when
dispersed in or dispersed
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
53
in water and then mixed with solutions with ionic strength corresponding to
Ca2+ equivalents of
about 0 to about 1 ppm, about 0 to about 10 ppm, about 0 to about 100 ppm,
about 0 to about 342
ppm, about 0 to about 500 ppm, about 0 to about 1000 ppm, about 0 to about
5000 ppm, about 0 to
about 8000 ppm, about 0 to about 10000 ppm, about 1 to about 10 ppm, about 1
to about 100 ppm,
about 1 to about 342 ppm, about 1 to about 500 ppm, about 1 to about 1000 ppm,
about 1 to about
5000 ppm, about 1 to about 8000 ppm, about 1 to about 10000 ppm, about 10 to
about 100 ppm,
about 10 to about 342 ppm, about 10 to about 500 ppm, about 10 to about 1000
ppm, about 10 to
about 5000 ppm, about 10 to about 8000 ppm, about 10 to about 10000 ppm, about
100 to about
342 ppm, about 100 to about 500 ppm, about 100 to about 1000 ppm, about 100 to
about 5000
ppm, about 100 to about 8000 ppm, about 100 to about 10000 ppm, about 342 to
about 500 ppm,
about 342 to about 1000 ppm, about 342 to about 5000 ppm, about 342 to about
8000 ppm, about
342 to about 10000 ppm, about 500 to about 1000 ppm, about 500 to about 5000
ppm, about 500
to about 8000 ppm, about 500 to about 10000 ppm, about 1000 to about 5000 ppm,
about 1000 to
about 8000 ppm, about 1000 to about 10000 ppm, about 5000 to about 8000 ppm,
about 5000 to
about 10000 ppm, about 8000 to about 10000 ppm.
Plant Health Applications
In some embodiments, the present disclosure provides formulations of triazoles
that have
both protective and curative activity. These formulations can be used as
protective fungicides,
curative fungicides, or as fungicides in both protective and curative
applications. These formulations
can be used at concentrations and use rates that correspond to any of the
values or ranges of values
noted above or in other portions of the Efficacy and Application Section.
In some embodiments, application of formulations of the present disclosure to
plants (e.g.,
crop plants) of the present disclosure results in a yield increase (e.g.,
increased crop yield). In some
embodiments, there is a yield increase compared to untreated crops. In some
embodiments, there is
an increase compared to crops that have been treated with a commercial
formulation of the same
active ingredient. In some embodiments, there is yield increase of about 2 to
about 100 %, e.g., 2 ¨ 3
%, 2 ¨ 5 %, 2- 10 %, 2-30 %, 2-50 %, 2-100 %, 5-7 %, 5-10 %, 5- 20 %, 5-30 %,
5-40%, 5- 50 %, 5- 60 %,
5- 70 %, 5- 80 %, 5- 90 %, 5- 100 %, 10¨ 20 %, 10-30 %, 10 ¨ 40 %, 10¨ 50%,
10¨ 60 %, 10 ¨ 70 %,
¨ 80 %, 10 ¨ 90 %, 20 ¨ 40 %, 20 ¨ 60 %, 20 ¨ 80 %, 20 ¨ 100 %, 30 ¨ 50 %, 30
¨ 60 %, 30 ¨ 80 %,
30 ¨ 100 %, 40 ¨ 60 %, 40 ¨ 80 %, 40 ¨ 100 %, 50 ¨ 80 %, 50 ¨ 100 %, 60 ¨ 80
%, 60 ¨ 100 %, 70 ¨ 90
%, 70 -100 % or 80 ¨ 100 %
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
54
In some embodiments, the use of the triazole formulations of the present
disclosure results
in a yield increase of about 2%, about 3%, about 4%, about 5%, about 6%, about
7%, about 10%,
about 20%, about 30%, about 40%, about 50 %, about 60%, about 70%, about 80%,
about 90% or
about 100%. In some embodiments, there is yield increase of greater than about
2 %, greater than
about 5%, greater than about 10 %, greater than about 20%, greater than about
30%, greater than
about 40%, greater than about 50 %, greater than about 60%, greater than about
70 %, greater
than about 80 %, greater than about 90% or greater than about 100 %. In some
embodiments, the
use of the triazole formulations of the present disclosure in plant health
applications results in a
yield increase of greater than about 10%, greater than about 20%, greater than
about 30%, about
40%, about 50%, about 60%, about 70%, about 80%, about 90% or about 100%. In
some
embodiments, there is an increase in yield of greater than about 10 %, greater
than about 20%,
greater than about 30%, greater than about 40%, greater than about 50%,
greater than about 60
%, greater than about 70%, greater than about 80 %, greater than about 90 % or
greater than about
100 %. Yield increases may be relative to untreated control plants (e.g.,
plants that have not been
treated with formulations of the present disclosure), or plants treated with
currently available
commercial products.
In some embodiments, inoculation of plants with formulations of the present
disclosure
provides an increased crop yield as described above, at an active ingredient
use rates that are lower
than the use rates listed on commercially available products of the same
active ingredient. In some
embodiments, the increased yield can correspond to any of the values or ranges
of values noted
above. In some embodiments, the increased yield is observed at an active
ingredient use rate that is
less than about 75%, less than 60%, less than 50%, less than 40%, less than
30%, less than 20% or
less than 10% of a rate listed on the label of commercially available
fungicide product of the same
active ingredient. In some embodiments, the increased yield is observed at an
active ingredient use
rate that is about 75%, about 60%, about 50 %, about 40%, about 30%, about 20%
or about 10%
of a rate listed on a label of a commercially available fungicide product of
the same active ingredient.
Labels of commercially available formulations often provide ranges of active
ingredient use rates to
inoculate plants. In some embodiments, inoculation of plants with a
formulation of the present
disclosure provides an increased crop yield at an active ingredient use rate
that is lower than the
minimum use rate of a range of use rates listed on the label of a commercially
available product. In
some embodiments inoculation of plants with a formulation of the present
disclosure provides an
increased crop yield at a use rate that is less than about 75 %, less than
about 60%, less than about
50%, less than about 40%, less than about 30%, less than about 20% or less
than about 10% of the
minimum use rate of a range of use rates listed on the label of a commercially
available product.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
Without wishing to be limited by any theory, in some embodiments, it is
thought that
increased yield is due enhanced plant health of plants treated with
formulations of the present
disclosure. As used herein, plant health refers to the overall condition of
the plant, including its size,
sturdiness, optimum maturity, consistency in growth pattern and reproductive
activity. As
mentioned above, optimizing and enhancing such factors is a goal of plant
breeders. As used herein,
increased or enhanced plant health can also refer to increased yield of one
sample or set of crops
(e.g., a crop field treated with fungicide) compared to another sample or set
of the same crops (e.g.,
an untreated crop field).
The enhancement of plant health by applications of triazole fungicides is
thought to be due
to a number of factors, as discussed above. These include combating hidden and
undiagnosed
diseases, as well as and triggering of plant growth regulator effects.
Additionally it is thought that
yield increases are a result of control of soil-borne disease or pests. In
some embodiments, the
triazole formulations of the present disclosure can be used to enhance plant
health at an active
ingredient use rate that is lower than the rate listed on the labels of
currently available commercial
curative fungicide products of the same active ingredient.
Without wishing to be limited by any theory, in some embodiments, it is
thought that the
formulations of the present disclosure can be used to enhance plant health at
an active ingredient
use rate that is lower than the rate listed on commercially available products
of the same active
ingredient due to their enhanced curative properties, ability to combat soil-
borne disease, hidden
disease and act as a more efficient plant growth regulator. Without wishing to
be limited by any
theory, it is though that in some embodiments, the enhanced properties are due
to enhanced foliar
penetration and/or translocation. Without wishing to be limited by any theory
it is thought that in
some embodiments, the formulations of the present disclosure are more
effective at combating
hidden disease because of their enhanced residual activity, which increases
the window of
opportunity for successful application timing.
Direct Soil & Seed Applications
In some embodiments, formulations of the current disclosure may be used to
control fungal
disease of plants (including seeds) by application to soil (inoculation of
soil). The formulations of the
current disclosure may be used to control fungal disease via application to
the soil in which a plant is
to be planted prior to planting (i.e., as pre-plant incorporated application).
In some embodiments,
the formulations of the present disclosure are used to control fungal disease
via inoculation of the
seed and soil at the time of seed planting (e.g., via an in-furrow application
or T-banded application).
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
56
The formulations of the current disclosure may also be applied to soil after
planting but prior to
emergence of the plant (i.e., as a pre-emergence application). In some
embodiments, soil is
inoculated with a formulation of the current disclosure via an aerosol spray
or pouring.
In some embodiments, the triazole formulations of the current disclosure may
be used to
control fungal diseases in the aforementioned applications at an active
ingredient use rate that is
lower than the use rate listed on the labels of commercially available
formulations of the same
active ingredient, as described above.
In some embodiments, the triazole formulations of the current disclosure can
be used to
control fungal disease when applied to seeds (e.g., via seed coating). In some
embodiments, the
formulations of the current disclosure are used to control fungal disease when
applied to seeds at an
active ingredient use rate that is less than the use rate of commercially
available formulations of the
same active ingredient. In some embodiments, a formulation of the present
disclosure is used to
control fungal diseases when applied to seeds at an active ingredient use rate
that is less than about
75%, less than about 60%, less than about 50%, less than about 40%, less than
about 30%, less
than about 20 % or less than about 10%, of a use rate listed on the label of a
currently available
commercial triazole product of the same active ingredient. In some
embodiments, a formulation of
the present disclosure are used to control fungal disease when applied to
seeds at an active
ingredient use rate that is about 75 %, about 60%, about 50%, about 40 %,
about 30%, about 20%
or about 10%, of a rate listed on the label of a currently available triazole
product of the same active
ingredient. In some embodiments, commercially available products provide
ranges of active
ingredient use rates to control fungal disease when applied to seeds.
Increased Re-Application Interval
Due to their enhanced curative and preventative properties, in some
embodiments, the
formulations of the present disclosure can be applied at greater time
intervals (i.e., the time
between distinct inoculations) than currently available formulations of the
same active ingredient.
Inoculation intervals can be found on the labels of currently available
commercial formulations and
are readily accessible and available. In some embodiments, the formulations of
the present
disclosure are applied at an interval that is 1 day, 2 days, 3 days, 4 days, 5
days, 6 days, 7 days, 8
days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days or 15 days longer
than commercial
formulations of the same active ingredient. In some cases, commercial
formulations are applied at
intervals that correspond to a range of intervals (e.g., 7-14 days). In such
cases, it is contemplated
that the formulations of the present disclosure can be applied at a range of
intervals whose shortest
endpoint, longest endpoint, or both shortest and longest endpoint are longer
than the
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
57
corresponding endpoints of currently available commercial formulations by any
of the values noted
above. In some embodiments, the triazole formulations of the present
disclosure can be applied at
an intervals of 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12
days, 13 days, 14 days, 15
days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days,
24 days, 25 days, 26
days, 27 days, 28 days, 29 days, 30 days, 31 days, 32 days, 33 days, 34 days,
35 days, 36 days, 37
days, 38 days, 39 days or 40 days. In some embodiments, the formulations of
the present disclosure
can be applied at a range from which the shortest and longest intervals
(endpoints) are taken from
any of the aforementioned values.
Specific Application (Plant & Fungi)
In some embodiments, the inoculation method is applied to individual plants or
fungi, or to
large groups of plants and fungi. In some embodiments, the formulation is
inoculated to the target
organism by means of dipping the target organism or part of the organism into
the dispersion
containing the formulation. In some embodiments, the formulation is inoculated
to the target
species (plant or fungi) by means of an aerosol spray. In some embodiments,
the formulation is
inoculated to the target species (plant) by spraying the dispersion directly
onto the leaves, stem,
bud, shoot or flowers of the plant. In some embodiments, the formulation is
inoculated to the target
species (plant) by pouring the dispersion directly onto the root zone of the
plant. In some
embodiments, the target organism (e.g., the plant on which fungus is to be
controlled or the fungus
is inoculated by means of dipping the plant or a part of parts of the target
plant into a dispersion of
active ingredients prepared as described above. Formulations of the current
invention can also be
applied in conjunction with irrigation systems and via water for irrigation.
The triazole formulations of the present disclosure can be used to control
fungal disease of a
variety of plants. In some embodiments, the plant is selected from the classes
fabaceaae,
brassicaceae, rosaceae, solanaceae, convolvulaceae, poaceae, annaranthaceae,
lanninaceae and
apiaceae .
In some embodiments, the plant is selected from plants that are grown for
turf, sod, seed
(e.g., grasses grown for seed), pasture or ornamentals. In some embodiments,
the plant is a crop,
including but not limited to cereals (e.g., wheat, maize, including field corn
and sweet corn, rice,
barley, oats etc.), soybean, cole crops, tobacco, oil crops, cotton, fruits
(e.g., pome fruits such as but
not limited to apples and pears), vine crops (e.g., cucurbits), legume
vegetables, bulb vegetables,
rapeseed, potatoes, greenhouse crops, and all other crops on which triazoles
are known to control
fungal disease. Lists of plants on which fungal diseases are controlled by
specific commercially
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
58
available triazole formulations can be found on their labels, which are
readily accessible and
available.
In some embodiments, the formulations of the current disclosure can be applied
to turf, sod,
seed, pasture or ornamental in combination with other pesticides (e.g.,
insecticides, fungicides,
herbicides). In particular, fungicides with a different mode of action from
the triazole may be used to
mitigate resistance development in targeted fungi. Exemplary additional
fungicides include
strobilurins (e.g., azoxystrobin, trifloxystrobin, pyraclostrobin,
fluoxastrobin), aromatic fungicides
(e.g., chlorothalonil), conazoles, dicarboxinnides, benzimidazoles,
carbannates, and others. For
example, to treat the turf anthracnose (E.g., Colletotrichum spp.,
Colletotrichum cerealis) fosetyl-Al,
several different strobilurins, nnancozeb, chlorothalonil, amongst others, can
be used in combination
with the disclosed formulations. Combination applications are not necessarily
limited to
combination of two active ingredients, but tertiary, quaternary and
combinations of five active
ingredients are more are possible with the formulations of the current
disclosure.
In some embodiments, the formulations of the current disclosure are used to
control fungal
diseases in turf, ornamental and non-crop applications (uses). Examples of
these applications can be
found on the labels of currently available triazole formulations, such as the
labels referenced in
other portions of the specification. Non-limiting examples of turf, ornamental
and non-crop
applications in which the formulations of the present disclosure can be used
include the control of
fungal diseases of turf (e.g., lawns and sod) in residential areas, athletic
fields, parks, and
recreational areas such as golf courses. Formulations of the present
disclosure may also be used to
control fungal diseases of ornamentals (e.g., shrubs, ornamental trees,
foliage plants etc.), including
ornamentals in or around any of the aforementioned areas, as well as in
greenhouses (e.g., those
used for growth of ornamentals). Examples of fungi that can be controlled in
turf, ornamental and
non-crop applications, include those listed as fungi turf, ornamental and non-
crop applications in any
other portion of the specification or in any of the labels of currently
available triazole products used
to control fungi in turf, ornamental and non-crop applications (such as the
those referenced in other
portions of the specification).
In some embodiments, the fungus to be controlled by the formulations of the
present
disclosure is selected from the classes asconnycota, basidionnycota,
deuteromycota,
blastocladionnycota, chytridionnycota, glomeronnycota and combinations
thereof.
Examples of fungal diseases that can be controlled with formulations of the
current
disclosure include but are not limited to various blights, spots and rusts,
rots, blasts and smuts and
combinations thereof.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
59
In some embodiments, the plant (e.g., crop) on which fungal disease can be
controlled by
formulations of the present disclosure may depend on, among other variables,
the active ingredient,
inclusion of other components into the formulation, and the particular
application. Common
commercial formulations frequently include labels and instructions describing
the compatibility of
actives, inclusion of additives, tank mixes with other products (e.g.,
surfactants) labeled fungi,
instructions and restrictions for particular applications and uses as well as
other information. Such
labels and instructions pertinent to the formulations of the present
disclosures and their application
are also contemplated as part of the present disclosures. Labels are readily
accessible from
manufacturers' websites, or via centralized internet databases such as
Greenbook
(http://www.greenbook.net/) or the Crop Data Management Systems website
(www.cdnns.net).
In some embodiments, the triazole of the present disclosure is difenconazole,
tebuconazole,
cyproconazole, epoxiconazole, flutriafol, ipconazole, nnetconazole, or
propiconazole.
Examples
Formulations
In the following formulation examples (1, 8- 10), particle sizes were measured
by DLS using a
Malvern Zetasizer ZS, except Examples 19 and 20.
Example 1: Preparation of a HSLS formulation of nanoparticles or aggregates of
nanoparticles of
polymer-associated difenoconazole via ball-milling [Nanoparticles derived from
p(MAA-co-S)
poly(methacrylic acid-co-styrene); 3:1 ratio of difenoconazole: nanoparticles]
Field Trial Code:
VCP-DFZ-01 in Example 3 ¨ Example 7 below and Figs. 1¨ 10.
136.7 g of technical grade difenoconazole (Pacific Agriscience, 95% purity),
43.33 g of
nanoparticles derived from poly(MAA-co-S) [MAA:S ratio = approximately 75:25
by weight], 14.44 g
of Geropon T-77, 21.67 g of Geropon TA/72, 2.18 g of AerosilTM 380 (fumed
silica), 7.22 g of AtloxTM
4913, 48.39 g of propylene glycol, 28.89 g Trans-10A (Trans-Chemco, Inc., 10 %
active anti-foam
silicone emulsion), 1.87 g of ProxelTM BD-20 (biocide, Industrial
Microbiostat, 19.3% active biocide
ingredient, Arch Chemicals Inc.) and 424.24 g of RO water were added to a
container and mixed for ¨
1 day with an overhead stirrer. After stirring, the mixture was distributed
into 30 mL vials. To each of
the vials were added stainless steel shots (20-30 mesh) to ¨1/3-1/2 of the
volume of the vial. Each of
the vials was secured to a vortex and shaken for 5 days. The sample was then
ball-milled in batches
according to the following procedure. To an 80 mL stainless steel milling jar
(EQ-MJ-3-80SS, MTI
Corporation, Richmond CA, USA) was added ¨40-50 mL of the mixture as well as
an approximately
equivalent volume of 2 mm stainless steel shots (shots were added until they
were just below the
surface of the liquid). The jar was sealed and milled on a desk top high speed
vibrating ball mill
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
(MSK-SF M-3, MTI Corporation, Richmond CA, USA) for 5 minutes, then cooled on
an ice bath for ¨5
minutes. Three additional milling/cooling cycles were performed (total of 4
cycles). The milled
formulation was filtered through a 150 p.m sieve. Viscosity: 22.5 cP at 24.1
C; assayed
difenoconazole content: 17% (w/w); Z-ave particle size (at 200 ppnn
difenoconazole in CIPAC D
water): 279 nm, polydispersity index: 0.26.
Example 2: Preparation of an HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated difenoconazole via ball-milling [Nanoparticles derived from
p(MAA-co-S)
poly(methacrylic acid-co-styrene); 3:1 ratio of difenoconazole: nanoparticles]
("VCP-05")
1321.9 g of technical grade difenoconazole (Pacific Agriscience, 95 % purity),
130 g of
Geropon T-77, 195 g of Geropon TA/72, 19.5 g of Aerosil 380 (fumed silica),
and 2586.5 g of RO
water were added to a container, mixed, and placed in an ice bath under
homogenization. The
homogenizer was run at 6000 rpm. With the homogenizer running at the
aforementioned speed,
the following were added in sequence: 435.5 g of propylene glycol; a slurry
containing 418.6 g of
nanoparticles derived from poly(MAA-co-S) [MAA:S ratio = approximately 75:25
by weight]; 16.25 g
of ProxelTM BD-20 (biocide, Industrial Microbiostat, 19.3% active biocide
ingredient, Arch Chemicals
Inc.); 26.0g Trans-10A (Trans-Chennco, Inc., 10% active anti-foam silicone
emulsion,); and 65 g of
AtIoxTM 4913. After the addition of these five components the homogenizer
speed was increased
to 8000 rpm, giving a tip speed of 2823 ft /nnin, The mixture was homogenized
at this speed until the
diameter of at least 99 % of the particles (D(v, 0.9)) was less than 80 p.m as
measured on a
Mastersizer, and the average particle size was between 20 ¨ 25 urn This was
accomplished after 80
minutes of homogenization.
After homogenization was complete, the mixture was transferred to a Dyno-M ill
(Model
KDL). The mixture was milled at 3000 rpm, resulting in a tip speed of 2,000
ft. / min. The mixture
was milled with beads having a diameter between 0.6 and 0.8 mm made of cerium
stabilized
zirconium oxide. The temperature of the milling chamber was maintained at 40
C or less. Milling
was completed when the average particle size was less than 0.3 p.m This was
achieved after 120
minutes of milling, when the average particle size measured 0.274 p.m.
Samples were taken and evaluated for particle size, viscosity, density, and an
HPLC assay of
active ingredient content. The average particle size of the final formulation
was 339 nnn, an increase
over the final measurement during mill due to possible post-milling
aggregation of the polymer-
associated active ingredient nanoparticles. The formulation had a density of
1.103 g/mL, a viscosity
of 71.9 cP at 25.1 C, a pH of 5.92 and contained 20.4% active ingredient. This
formulation is
commonly referred to as VCP-05 in the Examples below and in the Figures.
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
61
Formulation Testing
Several field trials were conducted to evaluate performance of difenoconazole
formulations
described in this disclosure, compare their performance to current
commercially available
formulations of difenoconazole (InspireTm), and compare their performance of
commonly used
fungicidal treatments for specific pest/crop applications. A variety of crops
and diseases were
tested, as described below.
Example 3: Treating Black Spot on Cabbage
Difenoconazole at three different application rates (75, 125 and 175 g
a.i./ha) was applied to
cabbage plants with Black Spot (pathogen: Alternaria brassicicola). Two
formulations were tested:
the first formulation was prepared according to Example 1, and the second was
a commercially-
available formulation (InspireTm). Both formulations were tank mixed with
water and a 0.5 vol % of a
non-ionic surfactant to the application rates for the trial. The non-ionic
surfactant selected was
InduceTm (alkylarylpolyoxyalkane ethers, fatty acids and dinnethyl
polysiloxane). Disease
development was evaluated 4 days after a second treatment, 5, 19, and 33 days
after a third
treatment. Both formulations demonstrated control across the range of
application rates. Rates of
disease control (averaged across the three application rates) are illustrated
in Figure 1, though
disease incidence among the untreated controls was low and the severity of
infection of the
untreated control as low as well.
Example 4: Treating Powdery Mildew on Cucurbit (Cantaloupes, Squash)
Difenoconazole at three different application rates (75, 125 and 175 g
a.i./ha) was applied to
cantaloupe plants with powdery mildew (pathogen: Golovinomyces cichoracearum).
Two
formulations were tested: the first formulation was prepared according to
Example 1 and the second
was a commercially-available formulation (InspireTm). Both formulations were
tank mixed with
water and a 0.1 vol % of a non-ionic surfactant to the application rates for
the trial. The non-ionic
surfactant selected was Dyne-AmicTM (methyl esters of C16-C18 fatty acids,
polyalkyleneoxide
modified polydinnethylsiloxane, alkylphenol ethoxylate). Disease development
was evaluated 6 and
11 days after a second treatment, 10 and 18 days after a third treatment. Both
formulations
demonstrated control across the range of application rates. Rates of disease
control are illustrated
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
62
in Figure 2 (control rates averaged across the three application rates) and
Figure 3 (control rates 18
days after third treatment for three application rates).
Difenoconazole at three different application rates (75, 125 and 175 g
a.i./ha) was applied to
squash plants with powdery mildew (pathogen: Podosphaera xanthii). Two
formulations were
tested, the first formulation was prepared according to Example 1 and a
commercially-available
formulation (InspireTm). Both formulations were tank mixed with water and a
0.25 vol % of a non-
ionic surfactant to the application rates for the trial. The non-ionic
surfactant selected was Dyne-
AmicTM. Disease development was evaluated 14 days after a second treatment.
Rates of disease
control 14 days after treatment are illustrated in Figure 4. Figure 5 shows
rates of control (incidence
in Figure 5A and severity Figure 5B) at an earlier evaluation time, 12 days
after second application.
Example 5: Treating Early and Late Leaf Spots on Peanut Plants
Difenoconazole at three different application rates (75, 125 and 175 g
a.i./ha) was applied to
peanuts with Peanut Leaf Spot (pathogen: Pseudocercospora personata). Two
formulations were
tested: the first formulation was prepared according to Example 1, and the
second was a
commercially-available formulation (InspireTm). Both formulations were tank
mixed with water and
a 1.0 vol % of a non-ionic surfactant to the application rates for the trial.
The non-ionic surfactant
selected was ScannerTM (3-oxapentane-1,5-diol, propane-1,2,3-triol,
alkylphenol ethoxylate,
polydimethylsiloxane) Disease development was evaluated 7, 19 and 27 days
after three treatments.
Both formulations demonstrated reduction in defoliation and enhancement based
on the use of the
non-ionic surfactant. See Figure 6. Untreated controls rates of defoliation
of: 69%, 7 days after
treatment; 95%, 19 days after treatment; and 100%, 27 days after treatment.
Efficacy was also
measured by yield rates (Figure 7). Formulations prepared according to Example
1 showed
improved reduction in defoliation and improved yield rates as compared to the
commercially
available formulation.
Example 6: Treating Frog-Eye Spot! Cercospora Leaf Spot on Soybeans
Difenoconazole at three different application rates (75, 125 and 175 g
a.i./ha) was applied to
soybeans with two foliar cercosporas, Frog-Eye Leaf Spot and Leaf Spot
(pathogens: Cercospora
sojina and Cercospora kikuchii, respectively). Two formulations were tested:
the first formulation
was prepared according to Example 1 and the second was a commercially-
available formulation
(InspireTm). Both formulations were tank mixed with water and a 1.0 vol % of a
non-ionic surfactant
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
63
to the application rates for the trial. The non-ionic surfactant selected was
InduceTM. Disease
development was evaluated 14 days after treatment. Both formulations
demonstrated control
across the range of application rates. Efficacy was measured in several ways,
including rates of
disease control (Figure 8) 14 days after application and yield rates (Figure
9). Rates of disease
control indicated equivalent control between commercially available
formulations and formulations
prepared according to Example 1.
Example 7: Treating Early Blight on Tomatoes
Difenoconazole at three different application rates (75, 125 and 175 g
a.i./ha) was applied to
tomatoes with Early Blight (pathogen: Alternaria tomatophila). Two
formulations were tested: the
first formulation was prepared according to Example 1 and the second was a
commercially-available
formulation (Inspire-fly). Both formulations were tank mixed with water and a
1.0 vol % of a non-
ionic surfactant to the application rates for the trial. The non-ionic
surfactant selected was First
ChoiceTv Spreader Sticker (alkylarylpolyoxyethylene oxides) Disease
development was evaluated 6
days after treatment. Both formulations demonstrated control across the range
of application rates.
Rates of disease control are illustrated in Figure 10.
Example 8: Second Field Trial Treating Powdery Mildew on Cucurbit (zucchini)
Difenoconazole at three different application rates (75, 125 and 175 g al./ha)
was applied to
zucchini plants with powdery mildew (pathogen: Golovinomyces cichoracearum).
Two formulations
were tested: the first formulation was prepared according to Example 2 and the
second was a
commercially-available emulsifiable concentrate formulation (InspireTm). Both
formulations were
tank mixed with water and a 0.5 vol % of a non-ionic surfactant to the
application rates for the trial.
The non-ionic surfactant selected was Dyne-AmicTM. Disease development was
evaluated 6 days
after the first, second and third treatments, and 14 days after a third
treatment. Both formulations
demonstrated control across the range of application rates. Rates of disease
control are illustrated
in Figure 11 (control rates averaged across the three application rates) and
Figure 12 (control rates
during the trial with the three application rates averages). Disease severity
for the untreated
controls was 50% at 6 days after first treatment, and reached 100%, 6 days
after the second
treatment. Disease severity for the untreated controls did not decrease from
100% at the evaluation
time points (6 days after the third treatment and 14 days after the third
treatment).
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
64
Example 9: Treating Sigatoka Leaf Spot on Bananas
Difenoconazole at three different application rates (250, 417 and 667 ppm) was
applied to
banana plants with Sigatoka Leaf Spot (pathogens: Mycosphaerella musicol /
Cercospora musae).
Three formulations were tested: the first formulation was prepared according
to Example 2; the
second was a commercially-available emulsifiable concentrate formulation
(Syngenta EC); and the
third a proprietary oil in water ("EW") formulation. All three formulations
were tank mixed with
water to the proper dilution (10 grams of active ingredient in 15 liters of
water) with no other
adjuvant or additive. Each plant in a test plot received 0.5 L of diluted
fungicide formulation per
treatment. Each plot contained 30 plants.
Disease development was evaluated 7 days after each of three treatments which
were each
applied 7 days apart. For the evaluation, disease index was calculated on the
following basis: 0%
indicates no disease present; 100 indicates 51% of the tested leaf surface was
covered with the pest
(Mycosphaerella musicol / Cercospora musae). Percent disease control was
calculated based on the
disease index of the untreated control at the specific time point in the
treatment regimen, at the
end of the treatment in this case. Zero percent disease control indicates that
the test being
evaluated demonstrated an equivalent disease index as the untreated control,
while 100 percent
disease control indicates that the pest was substantially eradicated from the
leaf surface.
All three formulations demonstrated control across the range of application
rates. Disease
index, as described above, for the different formulations applied at a
concentration of 667ppnn is
shown in Figure 13. Final disease control assessment is shown for each
formulation at different
application rates 14 days after the first treatment in Figure 14. Disease
control for the untreated
control, which serves as the basis for disease index and disease control
calculations, is also shown in
Figure 13. The formulation prepared according to Example 2 exhibited disease
control equivalent to
the commercial emulsifiable concentrate formation and superior to the oil-in-
water emulsion
formulation.
Example 10: Treating Peanut Leaf Spot on Peanuts
Difenoconazole at two different application rates (75, and 125 g a.i./ha) was
applied to
peanuts with Peanut Leaf Spot (pathogens: Cercospora arachidicola,
Mycosphaerella berkeleyi). Two
formulations were tested at these application rates. The first formulation was
prepared according to
Example 2 and the second was a commercially-available emulsifiable concentrate
formulation
(InspireTm). A third formulation was also tested. The third formulation used a
different triazole
active ingredient, tebuconazole (MuscleTm) at an application rate of 227 g
al/ha. The formulations
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
prepared according to Example 2 were tank mixed with water and a 0.25 vol % of
a non-ionic
surfactant to the application rates for the trial. The non-ionic surfactant
selected was InduceTM. The
other formulations were tank-mixed with water to the final application
concentration. The non-ionic
surfactant was eliminated because the two commercial formulations were
emulsifiable
concentrates, which generally demonstrate increased plant phytotoxicity when
mixed with
additional surfactants.
Disease development was evaluated 16, 29, 42, and 58 days after four
treatments. Disease
was evaluated on a scale of 1 ¨ 10, where 1 indicates no disease, a score of 4
indicates noticeable
defoliation and 10 indicates over 80% defoliation. Both difenoconazole
formulations demonstrated
reduction in defoliation and enhancement (averaged across application rates).
See Figure 15. The
difenoconazole formulation prepared according to Example 2 exhibited superior
disease control,
even at lower application rates, see Figure 16. Untreated controls
demonstrated defoliation rates of
over 80% at the end of the trial, 42 days after the fourth treatment.
Efficacy was also measured by yield rates (Figure 17). Formulations prepared
according to
Example 2 showed improved reduction in defoliation and improved yield rates as
compared to the
commercially available formulation. For comparison of yield, additional
fungicide formulations were
used in comparison (EchoTM (chlorothalonil), EchoTm/Provosem
(chlorothalonil/prothioconazole) as
well as an additional non-ionic-surfactant with the formulation of Example 2.
Example 11: Treating White Mold on Peanuts
Difenoconazole at two different application rates (75, and 125 g a.i./ha) was
applied to
peanuts with White Mold (pathogen: Athelia rolfsii). Two formulations were
tested, the first
formulation was prepared according to Example 2 ("VCP-05") and the second was
a commercially
available emulsifiable concentrate formulation (InspireTm). The formulation
prepared according to
Example 2 was tank mixed with water and/or one of two non-ionic surfactants
(1.0 vol % of non-
ionic surfactant) to the application rates for the trial. The non-ionic
surfactant selected was InduceTM
or Silwet-L77TM (trisiloxane ethoxylate). InspireTM has increased
phytotoxicity when mixed with a
non-ionic-surfactant, and was only tank-mixed with water. Four replicates for
each formulation
were performed, each contained two 32 foot long rows. Disease development was
evaluated at the
end of the field trial. Disease control is calculated based on the percent of
crop row feet infected
with the pathogen. All formulations demonstrated reduction in infection under
heavy disease
pressure, see Figure 18. Untreated controls demonstrated a rate of infection
over 80%.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
66
Efficacy was also measured by yield rates (see Figure 19). Formulations
prepared according
to Example 2 showed improved reduction in defoliation and improved yield rates
as compared to
the commercially available formulation. For comparison of yield rates,
additional formulations were
used in comparison (BravoTM (chlorothalonil), BravoTm/Provosem
(chlorothalonil/prothioconazole))
as well as an additional non-ionic-surfactant with the formulation of Example
2.
Example 12: Treating Dollar Spot on Creeping Bentgrass
Difenoconazole at three different application rates (0.25, 0.5, and 1 fluid
oz. of formulation
applied per 1000 square feet of treatment area) was applied to treat dollar
spot (pathogen:
Sclerotinia homoeocarpa) on creeping bentgrass. The difenoconazole formulation
was prepared
according to Example 2. Each formulation was tank-mixed with water and a non-
ionic surfactant,
PulseTM (polyether modified polysiloxane) to give the proper concentration of
difenoconazole for the
application rate and 0.5 vol % of the non-ionic surfactant. The tank-mix
solution was applied to four
replicates, each a 3' by 5' plot. Applications of difenoconazole were repeated
every 14 days and the
disease control rate was evaluated at several intervals (6 days after
treatment 1, 2 days after
treatment 2, 12 days after treatment 2, 8 days after treatment 3, 4, 14, 24
and 34 days after
treatment 4). Lesions in untreated controls were evaluated at the same times.
Disease control
rates are shown in Figure 20.
Disease control rates were calculated based on the number of lesions present
on untreated
control plots. Zero percent control indicates an equivalent number of lesions
in a particular test plot
as compared to the untreated control plot. Table 3 below shows the number of
lesions (i.e., disease
severity) for untreated controls used as the basis for the disease control
rate calculations.
Table 3
Evaluation Time (Days after Treatment) Number of Lesions
6 days after treatment 1 66
2 days after treatment 2 82
12 days after treatment 2 113
8 days after treatment 3 59
4 days after treatment 4 134
14 days after treatment 4 215
24 days after treatment 4 223
34 days after treatment 4 222
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
67
Example 13: Additional Comparison of Mixed Fungicides
(Difenoconazole/Azoxystrobin)
Formulations in Treating Dollar Spot on Creeping Bentgrass
As part of the same applications to treat dollar spot in creeping bentgrass,
the formulation
according to Example 2 was mixed with HeritageTm, a commercially available
formulation of the
fungicide azoxystrobin. This mixture was prepared to compare its agrochemical
performance to the
BriskwayTM formulation, which is a commercially available formulation of the
combination of
difenoconazole and azoxystrobin. The difenoconazole formulation of Example 2
was applied at a
rate of 0.2 fl. Oz. per 1000 sq. ft., and mixed with HeritageTm so that the
HeritageTm product was
applied at a rate of 0.6 fl. Oz. per 1000 sq. ft. BriskwayTM was applied at a
rate of 0.3 fl. Oz per 1000
sq. ft. The rates were selected so that the same amount of active ingredient
for each fungicide was
applied to the treatment area. As shown in Figure 21, the two formulations
provided similar rates of
disease control, which were, in turn comparable to the control rates shown in
Figure 20 and Example
11.
Example 14: Treating Anthracnose on Annual Bluegrass
Difenoconazole at three different application rates (0.25, 0.5, and 1 fluid
oz. of formulation
applied per 1000 square feet of treatment area) was applied to treat
anthracnose (pathogen:
Colletotrichum cerealis) on annual bluegrass. The difenoconazole formulation
was prepared
according to Example 2. Each formulation was tank-mixed with a non-ionic
surfactant, PulseTM.
Applications of difenoconazole were repeated every 14 days and the disease
control rate was
evaluated at several intervals (13 days after treatment 2, 9 days after
treatment 3, 7 days after
treatment 4, and 3 days after treatment 5). Disease control rates are shown in
Figure 22.
Ill: Additional Formulations
Example 15: Preparation of a solid formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated difenoconazole via spray drying from a common solvent (2:1
ratio of
difenoconazole: nanoparticles)
8 g of difenoconazole and 4 g of nanoparticles derived from p(MAA-co-BUMA)
[ratio of
MAA:BUMA = approximately 75:25 by weight] were dissolved in 80 mL of methanol
and spray dried
on a Yannato ADL-311S spray dryer equipped with a GAS-410 organic solvent
recovery unit. Outlet
temp: ¨96 C; Inlet temp.: ¨155 C; feed rate 17.5 nnLinnin; atomizing air:
0.05 MPa.
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
68
A similar procedure was used to prepare a solid formulation (2:1 ratio of
difenoconazole :
nanoparticles) from nanoparticles derived from poly(MAA-co-S) [ratio of MAA:S
= approximately
75:25].
Example 16: Preparation of a HSLS formulation from a solid formulation of
nanoparticles or
aggregates of nanoparticles of polymer-associated difenoconazole via ball-
milling [Nanoparticles
derived from p(MAA-co-BUMA); 2:1 ratio of difenoconazole: nanoparticles]
1.2 g of the solid formulation described in Example 15, 0.053 g of Geropon T-
77, 0.267 g of
Geropon TA/72, 0.053 g of Aerosil 380 (fumed silica), 0.357 g of propylene
glycol, 0.213 g of
Trans-10A (Trans-Chennco, Inc., 10 % active anti-foam silicone emulsion),
0.014 g of ProxelTm BD-20
(biocide, Industrial Microbiostat, 19.3% active biocide ingredient, Arch
Chemicals Inc.) and 3.176 g of
RO water were added to a vial along with stainless steel shots (20-30 mesh) in
an amount
corresponding to about 1/2 of the volume of the liquid. The vial was secured
to a vortex and shaken
for 3 days. When the resulting formulation was dispersed in RO water at 200
ppnn difenoconazole,
the Z-ave particle size was 772 nnn with a polydispersity of 0.24.
Example 17: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated difenoconazole via ball-milling [Nanoparticles derived from
p(MAA-co-BUMA)
poly(methacrylic acid-co-butylmethacrylate; 2:1 ratio of difenoconazole:
nanoparticles]
0.267 g of Geropon T-77, 1.33 g of Geropon TA/72, 0.267 g of Aerosil 380
(fumed silica),
1.79 g of propylene glycol, 1.07 g of Trans-10A (Trans-Chennco, Inc., 10%
active anti-foam silicone
emulsion), 0.069 g of Proxerm BD-20 (biocide, Industrial Microbiostat, 19.3%
active biocide
ingredient, Arch Chemicals Inc.) and 15.89 g of RO water were added to a vial
and mixed (pH 9). The
pH of was adjusted to 6.15 via the addition of about 0.3 mL of 4 M H2SO4 and
the resulting liquid was
mixed with 4.0 g of difenoconazole (technical grade) and 2.0 g of
nanoparticles derived from
p(MAA-co-BUMA) [ratio of MAA:BUMA = approximately 75:25 by weight. To a
stainless steel milling
jar (EQ-M1-3-80SS, MTI Corporation, Richmond CA, USA) were added the resulting
mixture and 2 mm
stainless steel shots (shots were added until they were just below the surface
of the liquid). The jar
was sealed and milled on a desk top high speed vibrating ball mill (MSK-SFM-3,
MTI Corporation,
Richmond CA, USA) for 6 minutes, then cooled on an ice bath for 5 minutes.
Three additional
milling/cooling cycles were performed (total of 4 cycles).
When the formulation was dispersed in RO water at 200 ppnn difenoconazole, the
Z-ave
particle size was found to be 484 nnn with a polydispersity of 0.47. The
formulation was stable upon
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
69
heating at 45 C or 54 C for four days, as well after four temperature cycles
between -10 C and 45 C
in a cycling chamber.
Example 18: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated difenoconazole via ball-milling [Nanoparticles derived from
p(MAA-co-EA); 5:1
ratio of difenoconazole : nanoparticles]
1.0 g of difenoconazole (technical grade), 0.20 g of nanoparticles derived
from p(MAA-co-
EA) [ratio of MAA:EA= approximately 75:25 by weight], 0.15 g of Morwet D-425,
0.025 g of Aerosil
380 (fumed silica), 0.335 g of propylene glycol, 0.20 g of Trans-10A (Trans-
Chennco, Inc., 10% active
anti-foam silicone emulsion), 0.013 g of ProxelTM BD-20 (biocide, Industrial
Microbiostat, 19.3%
active biocide ingredient, Arch Chemicals Inc.) and 2.98 g of RO water were
added to a glass vial
along with stainless steel shots (20-30 mesh) in an amount corresponding to
about 1/2 of the volume
of the mixture. The vial was secured to a vortex and shaken for about 3 days.
When the resulting
formulation was dispersed in RO waterat 200 ppm difenoconazole, the Z-ave
particle size was 528
nnn with a polydispersity of 0.3. 5 mg of Xanthan gum (0.10 g of a 5% aqueous
Xanthan gum solution
prepared form Kelzan M, CP Kelco U.S., Inc) was added to the formulation,
which was then secured
to a vortex and shaken for about 4 hours.
Example 19: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated Azoxystrobin/Difenoconazole (1.6 ratio) via ball milling
[Nanoparticles derived
from (PMAA-co-S; 75:25) slurry]
4.92g of technical grade azoxystrobin (Pacific Agrosciences), 3.08 g technical
grade
difenoconazole (Pacific Agriscience, 95 % purity), 10.88 g of a slurry
containing 14.7 wt%
nanoparticles derived from poly(MAA-co-S) [MAA:S ratio = approximately 75:25
by weight] in water,
0.40g Geropon T-77, 2.0g Geropon TA/72, 0.40g Atlox 4913, 2.68g propylene
glycol, 0.16g Trans-10A
solution, 0.02g ProxelTM BD-20 solution and 15.46 g deionized water were all
placed in an 80nnL glass
beaker and were mixed overnight with an overhead paddle stirrer at 300-500 rpm
for approximately
18 hours. This mixture was then placed in a stainless steel milling jar along
with stainless steel
milling balls (assorted sizes, 2nnnn-6nnnn) and was milled for 6 minutes, and
then cooled in an ice
bath. This process was repeated 2 more times. The resulting composition was
then filtered through
a 100 mesh sieve. The filtered sample was then divided into 2 separate 30 nnL
vials that contained
about 5-log of 0.6mm stainless steel milling beads. The vials were sealed and
were shaken on a
vortex shaker (400 rpm) for 72 hours. The final formulation had the following
properties: viscosity:
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
121 cP at 23.7 C; assayed difenoconazole content: 12.7 % (w/w), assayed
azoxystrobin content: 7.8
(w/w); Z-ave particle size (undiluted): 248 nm by Malvern Mastersizer.
Example 20: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated Azoxystrobin/Difenoconazole (1.6 ratio) via ball milling
[Nanoparticles derived
from (PMAA-co-S; 75:25) concentrated slurry]
4.92g of technical grade azoxystrobin, 3.08 g technical grade difenoconazole,
5.56g of a
slurry containing 28.8 wt% nanoparticles derived from poly(MAA-co-S) [MAA:S
ratio = approximately
75:25 by weight] in water, 0.40g Geropon T-77, 2.0g Geropon TA/72, 2.68g
propylene glycol, 0.16g
Trans-10A solution, 0.02g ProxelTM BD-20 solution, and 21.18 g deionized water
were all placed in an
80nnL glass beaker and were mixed overnight with an overhead paddle stirrer at
300-500 rpm for
approximately 18 hours. This mixture was them placed in a stainless steel
milling jar along with
stainless steel milling balls (assorted sizes, 2nnnn-6nnnn) and was milled for
6 minutes, and then
cooled in an ice bath. This process was repeated 2 more times. The resulting
composition was then
filtered through a 100 mesh sieve. The filtered sample was then divided into 2
separate 30 nnL vials
that contained about 5-log of 0.6mm stainless steel milling beads. The vials
were sealed and were
shaken on a vortex shaker (400 rpm) for 72 hours. The final formulation had
the following
properties: assayed difenoconazole content: 13.2 % (w/w), assayed azoxystrobin
content: 7.9 %
(w/w); Z-ave particle size (undiluted): 403 nm by Malvern Mastersizer.
Example 21: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated Azoxystrobin/Difenoconazole (1.24 ratio) via mixing
separate formulations
[Nanoparticles derived from (PMAA-co-S; 75:25) slurry]
A 15.3 wt% difenoconazole formulation was made according to Example 2.
Similarly, a 19.1
wt % azoxystrobin formulation was prepared by milling: 87.6 g of azoxystrobin
technical (Pacific
Agrosciences), 96.7 g of a slurry containing 29.3 wt% nanoparticles derived
from poly(MAA-co-S)
[MAA:S ratio = approximately 75:25 by weight] in water, 15.0 g of Geropon T-
77, 10.0g of Geropon
TA/72, 5.0g Atlox 4913, 32 mL propylene glycol, 20 nnL Trans 10-A antifoam
solution, 1 nnL ProxelTM
BD-10 solution and 230.6 nnL of water. The mixture was homogenized for 45 min
at 70,000 rpm,
then milled on an Eiger mill for 135 minutes at 4000 rpm. The final
azoxystrobin formulation had an
average particle size of 314.6 nm (diluted to 200 ppnn in CIPAC D water). The
polydispersity index
was 0.299. The assayed azoxystrobin content was 18.1% (w/w) and the viscosity
was 229.5 cPs at
25.3 C.
CA 02936966 2016-07-14
WO 2014/118753
PCT/1B2014/058719
71
25.02g of the azoxystrobin formulation described above, and 19.54g of the
difenoconazole
formulation described above were placed in a 50nnl_ Nalgene bottle. The bottle
was capped and
shaken on a vortex shaker at low setting for 12 hours. The mixed formulation
had an azoxystrobin-
difenoconazole ratio of 1.24.
Example 22: Preparation of an HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated tebuconazole via ball-milling [Nanoparticles derived from
p(MAA-co-S)
poly(methacrylic acid-co-styrene); 3:1 ratio of tebuconazole: nanoparticles]
8.358g of technical grade tebuconazole, 18.27g of a slurry containing 14.7 wt%
nanoparticles
derived from poly(MAA-co-S) [MAA:S ratio = approximately 75:25 by weight] in
water, 1.24g of
Geropon TA/72, 0.8167g of Geropon T-77, 0.4803g of Atlox 4913, 0.2331g of
Aerosil TM 380, 2.68g of
propylene glycol, 1.7301g of Trans-10A solution, 0.0989g of Proxel TM BD-20
solution and 6.7386 g
deionized water were all placed in a stainless steel milling jar along with
ceria coated milling balls
(assorted sizes, 0.6-0.8nnnn. The jar was sealed and was shaken for 5 minutes
by hand, followed by
milling for 5 minutes, and then cooled in an ice bath. The milling and cooling
steps were each
repeated 5 more times. The resulting composition was then filtered through a
100 mesh sieve.
Example 23: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated Azoxystrobin/Tebuconazole (1:1 ratio) via ball milling
[Nanoparticles derived
from (PMAA-co-S; 75:25) slurry]
4.1431g of technical grade tebuconazole, 4.1364g technical grade azoxystrobin,
18.1961g of
a slurry containing 14.7 wt% nanoparticles derived from poly(MAA-co-S) [MAA:S
ratio =
approximately 75:25 by weight] in water, 1.196g of Geropon TA/72, 0.8042g of
Geropon T-77,
0.2109g of Aerosil 380, 2.6299g of propylene glycol, 0.7973g of Trans-10A,
0.1073g of Proxel BD20
and 16.153 g of deionized water were all placed in a stainless steel milling
jar along with ceria coated
milling balls (assorted sizes, 0.6-0.8mm). The jar was sealed and was shaken
for 5 minutes by hand,
followed by milling for 5 minutes, and then cooled in an ice bath. The milling
and cooling steps were
each repeated 5 more times. The resulting composition was then filtered
through a 100 mesh sieve.
Example 24: Preparation of a HSLS formulation of nanoparticles or aggregates
of nanoparticles of
polymer-associated Azoxystrobin/Tebuconazole (1:1 ratio) via ball milling
[Nanoparticles derived
from (PMAA-co-S; 75:25) slurry]
4.1328g of technical grade tebuconazole, 4.122g technical grade azoxystrobin,
18.1634g of a
slurry containing 14.7 wt% nanoparticles derived from poly(MAA-co-S) [MAA:S
ratio = approximately
75:25 by weight] in water, 1.19966g of Geropon TA/72, 2.0122g of Calsoft AOS-
40, 0.2115g of
CA 02936966 2016-07-14
WO 2014/118753
PCT/IB2014/058719
72
Aerosil 380, 2.6622g of propylene glycol, 0.8077g of Trans-10A, 0.1031g of
Proxel TM BD-20 and
14.9119g of deionized water were all placed in a stainless steel milling jar
along with ceria coated
milling balls (assorted sizes, 0.6-0.8mm). The jar was sealed and was shaken
for 5 minutes by hand,
followed by milling for 5 minutes, and then cooled in an ice bath. The milling
and cooling steps were
each repeated 5 more times. The resulting composition was then filtered
through a 100 mesh sieve.
